Inhibitors of cxcr2

ABSTRACT

Compounds are provided as inhibitors of CXCR2, having the structure:

CROSS-REFERENCES TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.15/722,785 filed Oct. 2, 2017 which is a continuation of U.S. patentapplication Ser. No. 15/353,949 filed Nov. 17, 2016 (now U.S. Pat. No.9,809,581), which is an application claiming benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 62/257,529 filed Nov. 19,2015, each of which is herein incorporated by reference in its entirety.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH AND DEVELOPMENT

Not applicable

REFERENCE TO A “SEQUENCE LISTING,” A TABLE, OR A COMPUTER PROGRAMLISTING APPENDIX SUBMITTED ON A COMPACT DISK

Not applicable

BACKGROUND OF THE INVENTION

Chemokines are chemotactic cytokines that are released by a wide varietyof cells, to attract cells such as leukocytes (including macrophages,T-cells, eosinophils, basophils, neutrophils and myeloid-derivedsuppressor cells) and endothelial cells to sites of inflammation andtumor growth. There are two main classes of chemokines, theCXC-chemokines and the CC-chemokines. The class depends on whether thefirst two cysteines are adjacent (CC-chemokines), or are separated by asingle amino acid (CXC-chemokines). There are currently at least 17known CXC-chemokines, which include but are not limited to CXCL1 (GROα),CXCL2 (GROβ), CXCL3 (GROγ), CXCL4 (PF4), CXCL5 (ENA-78), CXCL6 (GCP-2,CXCL7 (NAP-2), CXCL8 (IL-8, NAP-I), CXCL9 (MIG) and CXCL10 (IP-10).There are currently at least 28 known CC chemokines, which include butare not limited to CCL2 (MCP-1), CCL3 (MIP-1α), CCL4 (MIP-1β), CCL5(RANTES), CCL7 (MCP-3), CCL8 (MCP-2), CCL-11 (eotaxin-1) and CCL20(MIP-3α). Individual members of the chemokine families are known to bebound by at least one chemokine receptor, with CXC-chemokines generallybound by members of the CXCR class of receptors, and CC-chemokines bymembers of the CCR class of receptors. For example, CXCL8/IL-8 is boundby the receptors CXCR1 and CXCR2.

Since CXC-chemokines often promote the accumulation and activation ofneutrophils, these chemokines are implicated in a wide range of acuteand chronic inflammatory disorders such as psoriasis, rheumatoidarthritis, radiation-induced fibrotic lung disease, autoimmune bullousdermatoses (AIBD), chronic obstructive pulmonary disease (COPD) andozone-induced airway inflammation (see, Baggiolini et al., FEBS Lett.307:97 (1992); Miller et al., Crit. Rev. Immunol. 12:17 (1992);Oppenheim et al., Annu. Rev. Immunol. 9: 617 (1991); Seitz et al., J.Clin. Invest. 87: 463 (1991); Miller et al., Ann. Rev. Respir. Dis.146:427 (1992); and Donnely et al., Lancet 341: 643 (1993), Fox &Haston, Radiation Oncology, 85:215 (2013), Hirose et al., J. Genet.Syndr. Genet. Ther. S3:005 (2013), Miller et al., Eur. J. Drug Metab.Pharmacokinet. 39:173 (2014), Lazaar et al., Br. J Clin. Pharmacol.,72:282 (2011)).

A subset of CXC chemokines, those which contain the ELR motif (ELR-CXC),have been implicated in the induction of tumor angiogenesis (new bloodvessel growth). These include the CXCR2 ligand chemokines CXCL-1, CXCL2,CXCL3, CXCL5 and (Strieter et al. JBC 270: 27348-27357 (1995)) SomeCXCR2 ligand ELR-CXC chemokines are exacerbating agents during ischemicstroke (Connell et al., Neurosci. Lett., 15:30111 (2015). All of thesechemokines are believed to exert their actions by binding to CXCR2.Thus, their angiogenic activity is due to their binding and activationof CXCR2 expressed on the surface of vascular endothelial cells (ECs) insurrounding vessels.

Many different types of tumors are known to produce ELR-CXC chemokines,and production of these chemokines correlates with a more aggressivephenotype (Inoue et al. Clin Cancer Res 6:2104-2119 (2000)) and poorprognosis (Yoneda et. al. J Nat Cancer Inst 90:447-454 (1998)). AsELR-CXC chemokines are potent chemotactic factors for EC chemotaxis,they probably induce chemotaxis of endothelial cells toward their siteof production in the tumor. This may be a critical step in the inductionof tumor angiogenesis. Inhibitors of CXCR2 will inhibit the angiogenicactivity of the ELR-CXC chemokines and therefore block the tumor growth.This anti-tumor activity has been demonstrated for antibodies to CXCL8(Arenberg et al. J Clin Invest 97:2792-2802 (1996)), ENA-78 (Arenberg etal. J Clin Invest 102:465-72 (1998)), and CXCL1 (Haghnegahdar et al. JLeukoc Biology 67:53-62 (2000)).

Many tumor cells express CXCR2 and tumor cells may thereby stimulatetheir own growth by secreting ELR-CXC chemokines. Thus, in addition towith decreasing angiogenesis within tumors, CXCR2 inhibitors maydirectly inhibit the growth of tumor cells.

CXCR2 is often expressed by myeloid-derived suppressor cells (MDSC)within the microenvironment of tumors. MDSC are implicated in thesuppression of tumor immune responses, and migration of MDSC in responseto CXCR2 ligand chemokines is most likely responsible for attractingthese cells into tumors. (see Marvel and Gabrilovich, J. Clin. Invest.13:1 (2015) and Mackall et al., Sci. Trans. Med. 6:237 (2014). Thus,CXCR2 inhibitors may reverse suppressive processes and thereby allowimmune cells to more effectively reject the tumor. In fact, blocking theactivation of CXC-chemokine receptors has proven useful as a combinationtherapy with checkpoint inhibitors in suppressing tumor growth,suggesting that CXCR2 blockade may also enhance tumor rejection incombination with other anti-tumor therapies, including but not limitedto vaccines or traditional cytotoxic chemotherapies (see Highfill etal., Science Translational Medicine, 6:237 (2014)).

Hence, the CXC-chemokine receptors represent promising targets for thedevelopment of novel anti-inflammatory and anti-tumor agents.

There remains a need for compounds that are capable of modulatingactivity at CXC-chemokine receptors. For example, conditions associatedwith an increase in IL-8 production (which is responsible for chemotaxisof neutrophil and T-cell subsets into the inflammatory site and growthof tumors) would benefit by compounds that are inhibitors of IL-8receptor binding.

BRIEF SUMMARY OF THE INVENTION

Provided herein, in one aspect, are compounds having formula (I),

In formula (I) above, and herein, R¹ and R² are each membersindependently selected from the group consisting of H, halogen, CN, C₁₋₄alkyl, C₁₋₄ alkoxy and C₁₋₄ haloalkyl; R^(3a) is a member selected fromthe group consisting of methyl, ethyl, propyl, isopropyl,trifluoromethyl, CH₂CF₃ and CF₂CF₃; R^(3b) is a member selected from thegroup consisting of H and D; R⁴ is a member selected from the groupconsisting of H, C₁₋₈ alkyl, —Y and C₁₋₄alkylene-Y; wherein Y is aryl orheteroaryl, and each R⁴ is optionally substituted with from one to foursubstituents selected from the group consisting of halogen, —CN,—CO₂R^(a), —CONR^(a)R^(b), —C(O)R^(a), —OC(O)NR^(a)R^(b),NR^(a)C(O)R^(b), NR^(a)C(O)₂R^(c), —NR^(a)C(O)NR^(a)R^(b), —NR^(a)R^(b),—OR^(a), —S(O)₂NR^(a)R^(b), —NR^(a)S(O)₂R^(b), and —R^(c), wherein eachR^(a) and R^(b) is independently selected from hydrogen, C₁₋₄ alkyl,C₁₋₄ hydroxyalkyl and C₁₋₄ haloalkyl, R^(c) is selected from C₁₋₄ alkyl,C₁₋₄ hydroxyalkyl and C₁₋₄ haloalkyl; R^(5a) and R^(5b) are each membersindependently selected from the group consisting of H, halogen, C₁₋₄alkyl, C₁₋₄ alkoxy and CN; R^(6a) and R^(6b) are each membersindependently selected from the group consisting of H, C₁₋₄ alkyl, C₁₋₄hydroxyalkyl and C₁₋₄ haloalkyl; or optionally R^(6a) and R^(6b) aretaken together to form oxo (═O); X is CH or N; or any salts, solvates,hydrates, N-oxides, tautomers or rotamers thereof.

The compounds provided herein are useful for selectively binding to, andinhibiting the activity of CXCR2, and treating diseases that aredependent, at least in part, on CXCR2 activity. Accordingly, the presentinvention provides in further aspects, compositions containing one ormore of the above-noted compounds in admixture with a pharmaceuticallyacceptable excipient.

In still another aspect, provided herein are methods for treatingvarious diseases, discussed further herein, comprising administering toa subject in need to such treatment a therapeutically effective amountof a compound of the above formula for a period of time sufficient totreat the disease.

In yet another aspect, provided herein are methods of diagnosing diseasein an individual. In these methods, the compounds provided herein areadministered in labeled form to a subject, followed by diagnosticimaging to determine the presence or absence of CXCR2. In a relatedaspect, a method of diagnosing disease is carried out by contacting atissue or blood sample with a labeled compound as provided herein anddetermining the presence, absence, or amount of CXCR2 in the sample.

In some embodiments, an amount of a chemotherapeutic agent or radiationis administered to the subject prior to, subsequent to or in combinationwith the compounds provided herein. In some embodiments, the amount issub-therapeutic when the chemotherapeutic agent or radiation isadministered alone.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-1J provide structures and biological activity for compoundsdescribed herein.

DETAILED DESCRIPTION OF THE INVENTION

Before the present invention is further described, it is to beunderstood that the invention is not limited to the particularembodiments set forth herein, and it is also to be understood that theterminology used herein is for the purpose of describing particularembodiments only, and is not intended to be limiting.

Where a range of values is provided, it is understood that eachintervening value, to the tenth of the unit of the lower limit unlessthe context clearly dictates otherwise, between the upper and lowerlimit of that range and any other stated or intervening value in thatstated range, is encompassed within the invention. The upper and lowerlimits of these smaller ranges may independently be included in thesmaller ranges, and are also encompassed within the invention, subjectto any specifically excluded limit in the stated range. Where the statedrange includes one or both of the limits, ranges excluding either orboth of those included limits are also included in the invention. Unlessdefined otherwise, all technical and scientific terms used herein havethe same meaning as commonly understood by one of ordinary skill in theart to which this invention belongs.

It must be noted that as used herein and in the appended claims, thesingular forms “a,” “an,” and “the” include plural referents unless thecontext clearly dictates otherwise. It is further noted that the claimsmay be drafted to exclude any optional element. As such, this statementis intended to serve as antecedent basis for use of such exclusiveterminology such as “solely,” “only” and the like in connection with therecitation of claim elements, or use of a “negative” limitation.

General

The present invention derives from the discovery that compounds offormula I act as potent and selective antagonists of the CXCR2 receptor.The compounds have in vivo anti-inflammatory activity and have superiorpharmacokinetic properties. Accordingly, the compounds provided hereinare useful in pharmaceutical compositions, methods for the treatment ofCXCR2-mediated diseases, and as controls in assays for theidentification of competitive CXCR2 antagonists.

Abbreviation and Definitions

The term “alkyl”, by itself or as part of another substituent, means,unless otherwise stated, a straight or branched chain hydrocarbonradical, having the number of carbon atoms designated (i.e. C₁₋₈ meansone to eight carbons). Examples of alkyl groups include methyl, ethyl,n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl,n-hexyl, n-heptyl, n-octyl, and the like. The term “alkenyl” refers toan unsaturated alkyl group having one or more double bonds. Similarly,the term “alkynyl” refers to an unsaturated alkyl group having one ormore triple bonds. Examples of such unsaturated alkyl groups includevinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl),2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl,3-butynyl, and the higher homologs and isomers. The term “cycloalkyl”refers to hydrocarbon rings having the indicated number of ring atoms(e.g., C₃₋₆cycloalkyl) and being fully saturated or having no more thanone double bond between ring vertices. “Cycloalkyl” is also meant torefer to bicyclic and polycyclic hydrocarbon rings such as, for example,bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, etc. The term“cycloalkenyl” refers to a cycloalkyl group having at least one doublebond between ring vertices. Examples of cycloalkenyl are cyclopentenyland cyclohexenyl. The term “spirocycloalkyl” refers to a cycloalkylgroup in which a single ring vertex is attached to two othernon-hydrogen portions of the molecule. A spirocycloalkyl substituent isone in which two carbon atoms of an alkylene chain (typically thetermini of the alkylene chain) are attached to the same carbon atom inthe remainder of the molecule. The term “heterocycloalkyl” refers to acycloalkyl group that contain from one to five heteroatoms selected fromN, O, and S, wherein the nitrogen and sulfur atoms are optionallyoxidized, and the nitrogen atom(s) are optionally quaternized. Theheterocycloalkyl may be a monocyclic, a bicyclic or a polycylic ringsystem. Non limiting examples of heterocycloalkyl groups includepyrrolidine, imidazolidine, pyrazolidine, butyrolactam, valerolactam,imidazolidinone, hydantoin, dioxolane, phthalimide, piperidine,1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S-oxide,thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrroline,thiopyran, pyrone, tetrahydrofuran, tetrhydrothiophene, quinuclidine,and the like. A heterocycloalkyl group can be attached to the remainderof the molecule through a ring carbon or a heteroatom.

The term “alkylene” by itself or as part of another substituent means adivalent radical derived from an alkane, as exemplified by—CH₂CH₂CH₂CH₂—. Typically, an alkyl (or alkylene) group will have from 1to 24 carbon atoms, with those groups having 10 or fewer carbon atomsbeing preferred in the present invention. A “lower alkyl” or “loweralkylene” is a shorter chain alkyl or alkylene group, generally havingfour or fewer carbon atoms. Similarly, “alkenylene” and “alkynylene”refer to the unsaturated forms of “alkylene” having double or triplebonds, respectively.

As used herein, a wavy line, “

”, that intersects a single, double or triple bond in any chemicalstructure depicted herein, represent the point attachment of the single,double, or triple bond to the remainder of the molecule. Additionally, abond extending to the center of a ring (e.g., a phenyl ring) is meant toindicate attachment at any of the available ring vertices. One of skillin the art will understand that multiple substituents shown as beingattached to a ring will occupy ring vertices that provide stablecompounds and are otherwise sterically compatible.

The terms “alkoxy,” “alkylamino” and “alkylthio” (or thioalkoxy) areused in their conventional sense, and refer to those alkyl groupsattached to the remainder of the molecule via an oxygen atom, an aminogroup, or a sulfur atom, respectively. Additionally, for dialkylaminogroups, the alkyl portions can be the same or different and can also becombined to form a 3-7 membered ring with the nitrogen atom to whicheach is attached. Accordingly, a group represented as —NR^(a)R^(b) ismeant to include piperidinyl, pyrrolidinyl, morpholinyl, azetidinyl andthe like.

The terms “halo” or “halogen,” by themselves or as part of anothersubstituent, mean, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom. Additionally, terms such as “haloalkyl,” aremeant to include monohaloalkyl and polyhaloalkyl. For example, the term“C₁₋₄ haloalkyl” is mean to include trifluoromethyl,2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

The term “aryl” means, unless otherwise stated, a polyunsaturated,typically aromatic, hydrocarbon group which can be a single ring ormultiple rings (up to three rings) which are fused together or linkedcovalently. The term “heteroaryl” refers to aryl groups (or rings) thatcontain from one to five heteroatoms selected from N, O, and S, whereinthe nitrogen and sulfur atoms are optionally oxidized, and the nitrogenatom(s) are optionally quaternized. A heteroaryl group can be attachedto the remainder of the molecule through a heteroatom. Non-limitingexamples of aryl groups include phenyl, naphthyl and biphenyl, whilenon-limiting examples of heteroaryl groups include pyridyl, pyridazinyl,pyrazinyl, pyrimindinyl, triazinyl, quinolinyl, quinoxalinyl,quinazolinyl, cinnolinyl, phthalazinyl, benzotriazinyl, purinyl,benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzisoxazolyl,isobenzofuryl, isoindolyl, indolizinyl, benzotriazinyl, thienopyridinyl,thienopyrimidinyl, pyrazolopyrimidinyl, imidazopyridines,benzothiaxolyl, benzofuranyl, benzothienyl, indolyl, quinolyl,isoquinolyl, isothiazolyl, pyrazolyl, indazolyl, pteridinyl, imidazolyl,triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrrolyl,thiazolyl, furyl, thienyl and the like. Substituents for each of theabove noted aryl and heteroaryl ring systems are selected from the groupof acceptable substituents described below.

The term “arylalkyl” is meant to include those radicals in which an arylgroup is attached to an alkyl group (e.g., benzyl, phenethyl, and thelike). Similarly, the term “heteroaryl-alkyl”is meant to include thoseradicals in which a heteroaryl group is attached to an alkyl group(e.g., pyridylmethyl, thiazolylethyl, and the like).

The above terms (e.g., “alkyl,” “aryl” and “heteroaryl”), in someembodiments, will include both substituted and unsubstituted forms ofthe indicated radical. Preferred substituents for each type of radicalare provided below.

Substituents for the alkyl radicals (including those groups oftenreferred to as alkylene, alkenyl, alkynyl and cycloalkyl) can be avariety of groups selected from: -halogen, —OR′, —NR′R″, —SR′,—SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO₂R′, —CONR′R″, —OC(O)NR′R″,—NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)₂R′, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH,—NH—C(NH₂)═NR′, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NR'S(O)₂R″, —CN and—NO₂ in a number ranging from zero to (2 m′+1), where m′ is the totalnumber of carbon atoms in such radical. R′, R″ and R′″ eachindependently refer to hydrogen, unsubstituted C₁₋₈ alkyl, unsubstitutedaryl, aryl substituted with 1-3 halogens, unsubstituted C₁₋₈ alkyl, C₁₋₈alkoxy or C₁₋₈ thioalkoxy groups, or unsubstituted aryl-C₁₋₄ alkylgroups. When R′ and R″ are attached to the same nitrogen atom, they canbe combined with the nitrogen atom to form a 3-, 4-, 5-, 6-, or7-membered ring. For example, —NR′R″ is meant to include 1-pyrrolidinyland 4-morpholinyl.

Similarly, substituents for the aryl and heteroaryl groups are variedand are generally selected from: -halogen, —OR′, —OC(O)R′, —NR′R″, —SR′,—R′, —CN, —NO₂, —CO₂R′, —CONR′R″, —C(O)R′, —OC(O)NR′R″, —NR″C(O)R′,—NR″C(O)₂R′, —NR′—C(O)NR″R′″, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH,—NH—C(NH₂)═NR′, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NR'S(O)₂R″, —N₃,perfluoro(C₁-C₄)alkoxy, and perfluoro(C₁-C₄)alkyl, in a number rangingfrom zero to the total number of open valences on the aromatic ringsystem; and where R′, R″ and R′″ are independently selected fromhydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl, C₂₋₈ alkenyl,C₂₋₈ alkynyl, unsubstituted aryl and heteroaryl, (unsubstitutedaryl)-C₁₋₄ alkyl, and unsubstituted aryloxy-C₁₋₄ alkyl. Other suitablesubstituents include each of the above aryl substituents attached to aring atom by an alkylene tether of from 1-4 carbon atoms.

Two of the substituents on adjacent atoms of the aryl or heteroaryl ringmay optionally be replaced with a substituent of the formula-T-C(O)—(CH₂)_(q)—U—, wherein T and U are independently —NH—, —O—, —CH₂—or a single bond, and q is an integer of from 0 to 2. Alternatively, twoof the substituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula-A-(CH₂)_(r)—B—, wherein A and B are independently —CH₂—, —O—, —NH—,—S—, —S(O)—, —S(O)₂—, —S(O)₂NR′— or a single bond, and r is an integerof from 1 to 3. One of the single bonds of the new ring so formed mayoptionally be replaced with a double bond. Alternatively, two of thesubstituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula—(CH₂)_(s)—X—(CH₂)_(t)—, where s and t are independently integers offrom 0 to 3, and X is —O—, —NR′—, —S—, —S(O)—, —S(O)₂—, or —S(O)₂NR′—.The substituent R′ in —NR′— and —S(O)₂NR′— is selected from hydrogen orunsubstituted C₁₋₆ alkyl.

As used herein, the term “heteroatom” is meant to include oxygen (O),nitrogen (N), sulfur (S) and silicon (Si).

When a variable (e.g., R¹ or R^(a)) occurs more than one time in anyconstituent, its definition on each occurrence is independent of itsdefinition at every other occurrence. Additionally, combinations ofsubstituents and/or variables are permissible only if such combinationsresult in stable compounds.

The term “pharmaceutically acceptable salts” is meant to include saltsof the active compounds which are prepared with relatively nontoxicacids or bases, depending on the particular substituents found on thecompounds described herein. When compounds provided herein containrelatively acidic functionalities, base addition salts can be obtainedby contacting the neutral form of such compounds with a sufficientamount of the desired base, either neat or in a suitable inert solvent.Examples of salts derived from pharmaceutically-acceptable inorganicbases include aluminum, ammonium, calcium, copper, ferric, ferrous,lithium, magnesium, manganic, manganous, potassium, sodium, zinc and thelike. Salts derived from pharmaceutically-acceptable organic basesinclude salts of primary, secondary and tertiary amines, includingsubstituted amines, cyclic amines, naturally-occurring amines and thelike, such as arginine, betaine, caffeine, choline,N,N′-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol,2-dimethylaminoethanol, ethanolamine, ethylenediamine,N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine,hydrabamine, isopropylamine, lysine, methylglucamine, morpholine,piperazine, piperidine, polyamine resins, procaine, purines,theobromine, triethylamine, trimethylamine, tripropylamine, tromethamineand the like. When compounds provided herein contain relatively basicfunctionalities, acid addition salts can be obtained by contacting theneutral form of such compounds with a sufficient amount of the desiredacid, either neat or in a suitable inert solvent. Examples ofpharmaceutically acceptable acid addition salts include those derivedfrom inorganic acids like hydrochloric, hydrobromic, nitric, carbonic,monohydrogencarbonic, phosphoric, monohydrogenphosphoric,dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, orphosphorous acids and the like, as well as the salts derived fromrelatively nontoxic organic acids like acetic, propionic, isobutyric,malonic, benzoic, succinic, suberic, fumaric, mandelic, phthalic,benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, andthe like. Also included are salts of amino acids such as arginate andthe like, and salts of organic acids like glucuronic or galactunoricacids and the like (see, for example, Berge, S. M., et al,“Pharmaceutical Salts”, Journal of Pharmaceutical Science, 1977, 66,1-19). Certain specific compounds provided herein contain both basic andacidic functionalities that allow the compounds to be converted intoeither base or acid addition salts.

The neutral forms of the compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the present invention.

In addition to salt forms, the present invention provides compoundswhich are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds provided herein.Additionally, prodrugs can be converted to the compounds provided hereinby chemical or biochemical methods in an ex vivo environment. Forexample, prodrugs can be slowly converted to the compounds providedherein when placed in a transdermal patch reservoir with a suitableenzyme or chemical reagent.

Certain compounds provided herein can exist in unsolvated forms as wellas solvated forms, including hydrated forms. In general, the solvatedforms are equivalent to unsolvated forms and are intended to beencompassed within the scope of the present invention. Certain compoundsprovided herein may exist in multiple crystalline or amorphous forms. Ingeneral, all physical forms are equivalent for the uses contemplated bythe present invention and are intended to be within the scope of thepresent invention.

Certain compounds provided herein possess asymmetric carbon atoms(optical centers) or double bonds; the racemates, diastereomers,geometric isomers, regioisomers and individual isomers (e.g., separateenantiomers) are all intended to be encompassed within the scope of thepresent invention. In some embodiments, the compounds of the inventionare present in an enantiomerically enriched form, wherein the amount ofenantiomeric excess for a particular enantiomer is calculated by knownmethods. The preparation of enantiomerically enriched forms is also wellknown in the art and can be accomplished using, for example, chiralresolution via chromatography or via chiral salt formation. When aparticular stereochemical depiction is shown herein, it is meant torefer to that form of the compound in the stereochemistry is as shownand is substantially free of the other isomer. ‘Substantially free of’another isomer indicates at least an 80/20 ratio of the two isomers,more preferably 90/10, or 95/5 or more. In some embodiments, one of theisomers will be present in an amount of at least 99%. Additionally,different conformers are contemplated by the present invention, as wellas distinct rotamers. Conformers are conformational isomers that candiffer by rotations about one or more 6 bonds. Rotamers are conformersthat differ by rotation about only a single 6 bond. Still further, thecompounds provided herein may also contain unnatural proportions ofatomic isotopes at one or more of the atoms that constitute suchcompounds. Accordingly, in some embodiments, the compounds of theinvention are present in isotopically enriched form. Unnaturalproportions of an isotope may be defined as ranging from the amountfound in nature to an amount consisting of 100% of the atom in question.For example, the compounds may incorporate radioactive isotopes, such asfor example tritium (³H), iodine-125 (¹²⁵I) or carbon-14 (¹⁴C), ornon-radioactive isotopes, such as deuterium (²H) or carbon-13 (¹³C).Such isotopic variations can provide additional utilities to thosedescribed elsewhere with this application. For instance, isotopicvariants of the compounds of the invention may find additional utility,including but not limited to, as diagnostic and/or imaging reagents, oras cytotoxic/radiotoxic therapeutic agents. Additionally, isotopicvariants of the compounds of the invention can have alteredpharmacokinetic and pharmacodynamic characteristics which can contributeto enhanced safety, tolerability or efficacy during treatment. Allisotopic variations of the compounds provided herein, whetherradioactive or not, are intended to be encompassed within the scope ofthe present invention. Certain compounds provided herein are shown inone tautomeric form (e.g., a pyridone form), which is understood by oneof skill in the art to include the shown form as well as the othertautomeric form (e.g., a hydroxyl-pyridine).

“CXCR2” refers to CXC Chemokine Receptor 2, also known as CD128, IL8RBand L8 receptor type B, whose gene is encoded on human chromosome 2q35and is a known receptor for CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL7 andCXCL8 (see Murphy, P. M., Annu. Rev. Immunol. 12:593 (1994) and Zlotnik& Yoshie, Immunity, 12:127 (2000)).

The terms “patient” or “subject” are used interchangeably to refer to ahuman or a non-human animal (e.g., a mammal).

The terms “administration”, “administer” and the like, as they apply to,for example, a subject, cell, tissue, organ, or biological fluid, referto contact of, for example, an inhibitor of CXCR2, a pharmaceuticalcomposition comprising same, or a diagnostic agent to the subject, cell,tissue, organ, or biological fluid. In the context of a cell,administration includes contact (e.g., in vitro or ex vivo) of a reagentto the cell, as well as contact of a reagent to a fluid, where the fluidis in contact with the cell.

The terms “treat”, “treating”, treatment” and the like refer to a courseof action (such as administering an inhibitor of CXCR2 or apharmaceutical composition comprising same) initiated after a disease,disorder or condition, or a symptom thereof, has been diagnosed,observed, and the like so as to eliminate, reduce, suppress, mitigate,or ameliorate, either temporarily or permanently, at least one of theunderlying causes of a disease, disorder, or condition afflicting asubject, or at least one of the symptoms associated with a disease,disorder, condition afflicting a subject. Thus, treatment includesinhibiting (e.g., arresting the development or further development ofthe disease, disorder or condition or clinical symptoms associationtherewith) an active disease.

The term “in need of treatment” as used herein refers to a judgment madeby a physician or other caregiver that a subject requires or willbenefit from treatment. This judgment is made based on a variety offactors that are in the realm of the physician's or caregiver'sexpertise.

The terms “prevent”, “preventing”, “prevention” and the like refer to acourse of action (such as administering a CXCR2 inhibitor or apharmaceutical composition comprising same) initiated in a manner (e.g.,prior to the onset of a disease, disorder, condition or symptom thereof)so as to prevent, suppress, inhibit or reduce, either temporarily orpermanently, a subject's risk of developing a disease, disorder,condition or the like (as determined by, for example, the absence ofclinical symptoms) or delaying the onset thereof, generally in thecontext of a subject predisposed to having a particular disease,disorder or condition. In certain instances, the terms also refer toslowing the progression of the disease, disorder or condition orinhibiting progression thereof to a harmful or otherwise undesiredstate.

The term “in need of prevention” as used herein refers to a judgmentmade by a physician or other caregiver that a subject requires or willbenefit from preventative care. This judgment is made based on a varietyof factors that are in the realm of a physician's or caregiver'sexpertise.

The phrase “therapeutically effective amount” refers to theadministration of an agent to a subject, either alone or as part of apharmaceutical composition and either in a single dose or as part of aseries of doses, in an amount capable of having any detectable, positiveeffect on any symptom, aspect, or characteristic of a disease, disorderor condition when administered to the subject. The therapeuticallyeffective amount can be ascertained by measuring relevant physiologicaleffects, and it can be adjusted in connection with the dosing regimenand diagnostic analysis of the subject's condition, and the like. By wayof example, measurement of the serum level of an CXCR2 inhibitor (or,e.g., a metabolite thereof) at a particular time post-administration maybe indicative of whether a therapeutically effective amount has beenused.

The phrase “in a sufficient amount to effect a change” means that thereis a detectable difference between a level of an indicator measuredbefore (e.g., a baseline level) and after administration of a particulartherapy. Indicators include any objective parameter (e.g., serumconcentration) or subjective parameter (e.g., a subject's feeling ofwell-being).

The term “small molecules” refers to chemical compounds having amolecular weight that is less than about 10 kDa, less than about 2 kDa,or less than about 1 kDa. Small molecules include, but are not limitedto, inorganic molecules, organic molecules, organic molecules containingan inorganic component, molecules comprising a radioactive atom, andsynthetic molecules. Therapeutically, a small molecule may be morepermeable to cells, less susceptible to degradation, and less likely toelicit an immune response than large molecules.

The terms “inhibitors” and “antagonists”, or “activators” and “agonists”refer to inhibitory or activating molecules, respectively, for example,for the activation of, e.g., a ligand, receptor, cofactor, gene, cell,tissue, or organ. Inhibitors are molecules that decrease, block,prevent, delay activation, inactivate, desensitize, or down-regulate,e.g., a gene, protein, ligand, receptor, or cell. Activators aremolecules that increase, activate, facilitate, enhance activation,sensitize, or up-regulate, e.g., a gene, protein, ligand, receptor, orcell. An inhibitor may also be defined as a molecule that reduces,blocks, or inactivates a constitutive activity. An “agonist” is amolecule that interacts with a target to cause or promote an increase inthe activation of the target. An “antagonist” is a molecule that opposesthe action(s) of an agonist. An antagonist prevents, reduces, inhibits,or neutralizes the activity of an agonist, and an antagonist can alsoprevent, inhibit, or reduce constitutive activity of a target, e.g., atarget receptor, even where there is no identified agonist.

The terms “modulate”, “modulation” and the like refer to the ability ofa molecule (e.g., an activator or an inhibitor) to increase or decreasethe function or activity of CXCR2, either directly or indirectly. Amodulator may act alone, or it may use a cofactor, e.g., a protein,metal ion, or small molecule. Examples of modulators include smallmolecule compounds and other bioorganic molecules. Numerous libraries ofsmall molecule compounds (e.g., combinatorial libraries) arecommercially available and can serve as a starting point for identifyinga modulator. The skilled artisan is able to develop one or more assays(e.g., biochemical or cell-based assays) in which such compoundlibraries can be screened in order to identify one or more compoundshaving the desired properties; thereafter, the skilled medicinal chemistis able to optimize such one or more compounds by, for example,synthesizing and evaluating analogs and derivatives thereof. Syntheticand/or molecular modeling studies can also be utilized in theidentification of an Activator.

The “activity” of a molecule may describe or refer to the binding of themolecule to a ligand or to a receptor; to catalytic activity; to theability to stimulate gene expression or cell signaling, differentiation,or maturation; to antigenic activity; to the modulation of activities ofother molecules; and the like. The term “proliferative activity”encompasses an activity that promotes, that is necessary for, or that isspecifically associated with, for example, normal cell division, as wellas cancer, tumors, dysplasia, cell transformation, metastasis, andangiogenesis.

Embodiments of the Invention

A. Compounds

Provided herein, in one aspect, are compounds having formula (I),

In formula (I) above, and herein, R¹ and R² are each membersindependently selected from the group consisting of H, halogen, CN, C₁₋₄alkyl, C₁₋₄ alkoxy and C₁₋₄ haloalkyl; R^(3a) is a member selected fromthe group consisting of methyl, ethyl, propyl, isopropyl,trifluoromethyl, CH₂CF₃ and CF₂CF₃; R^(3b) is a member selected from thegroup consisting of H and D; R⁴ is a member selected from the groupconsisting of H, C₁₋₈ alkyl, —Y and C₁₋₄alkylene-Y; wherein Y is aryl orheteroaryl, and each R⁴ is optionally substituted with from one to foursubstituents selected from the group consisting of halogen, —CN,—CO₂R^(a), —CONR^(a)R^(b), —C(O)R^(a), —OC(O)NR^(a)R^(b),NR^(a)C(O)R^(b), —NR^(a)C(O)₂R^(c), —NR^(a)C(O)NR^(a)R^(b),—NR^(a)R^(b), —OR^(a), —S(O)₂NR^(a)R^(b), —NR^(a)S(O)₂R^(b), and —R^(c),wherein each R^(a) and R^(b) is independently selected from hydrogen,C₁₋₄ alkyl, C₁₋₄ hydroxyalkyl and C₁₋₄ haloalkyl, R^(c) is selected fromC₁₋₄ alkyl, C₁₋₄ hydroxyalkyl and C₁₋₄ haloalkyl; R^(5a) and R^(5b) areeach members independently selected from the group consisting of H,halogen, C₁₋₄ alkyl, C₁₋₄ alkoxy and CN; R^(6a) and R^(6b) are eachmembers independently selected from the group consisting of H, C₁₋₄alkyl, C₁₋₄ hydroxyalkyl and C₁₋₄ haloalkyl; or optionally R^(6a) andR^(6b) are taken together to form oxo (═O); X is CH or N; or any salts,solvates, hydrates, N-oxides, tautomers or rotamers thereof.

Selected embodiments are those in which (1) R¹ is selected from H, Cland CH₃; or (2) R² is H; or (3) R^(3a) is ethyl or isopropyl; or (4a)R^(3b) is H; or (4b) R^(3b) is D; or (5) X is CH; or (6) each of R^(5a)and R^(5b) are independently selected from H, Cl and F; or (7) each ofR^(6a) and R^(6b) are independently selected from H and C₁₋₄ alkyl; or(8) R⁴ is C₁₋₈ alkyl, optionally substituted with -halogen, —CN,—CO₂R^(a), —CONR^(a)R^(b), —C(O)R^(a), —OC(O)NR^(a)R^(b),—NR^(a)C(O)R^(b), —NR^(a)C(O)₂R^(c), —NR^(a)C(O)NR^(a)R^(b),—NR^(a)R^(b), —OR^(a), —S(O)₂NR^(a)R^(b), and —NR^(a)S(O)₂R^(b).Combinations of two or more, three or more, four or more, or five ormore of embodiments (1) through (8) are also contemplated as furtherselected embodiments.

In further selected embodiments, compounds are provided having formula(Ia):

wherein R¹ is selected from the group consisting of Cl and CH₃; R^(3b)is selected from the group consisting of H and D; R⁴ is a memberselected from the group consisting of H and C₁₋₈ alkyl, wherein the C₁₋₈alkyl is optionally substituted with —CONR^(a)R^(b), —OC(O)NR^(a)R^(b),—NR^(a)C(O)R^(b), —NR^(a)C(O)₂R^(c), —NR^(a)R^(b), or —OR^(a), whereineach R^(a) and R^(b) is independently selected from hydrogen, C₁₋₄alkyl, C₁₋₄ hydroxyalkyl and C₁₋₄ haloalkyl, and R^(c) is selected fromC₁₋₄ alkyl, C₁₋₄ hydroxyalkyl and C₁₋₄ haloalkyl; R^(5a) and R^(5b) areeach members independently selected from the group consisting of H, F,Cl and CH₃; R^(6a) and R^(6b) are each members independently selectedfrom the group consisting of H, C₁₋₄ alkyl, C₁₋₄ hydroxyalkyl and C₁₋₄haloalkyl; or optionally R^(6a) and R^(6b) are taken together to formoxo (═O); or any salts, solvates, hydrates, N-oxides or rotamersthereof.

Within formula (Ia), further selected embodiments are those in whichR^(3b) is H; R⁴ is H or CH₃; R^(5a) is H, F or Cl; R^(5b) is H, F, Cl;R^(6a) and R^(6b) are independently selected from the group consistingof H and CH₃, or are taken together to form oxo (═O).

Within formula (Ia), still other selected embodiments are those in whichR^(3b) is D; R⁴ is H or CH₃; R^(5a) is H, F or Cl; R^(5b) is H, F, Cl;R^(6a) and R^(6b) are independently selected from the group consistingof H and CH₃, or are taken together to form oxo (═O).

In other selected embodiments, compounds are provided having formula(Ib):

wherein R¹ is selected from the group consisting of Cl and CH₃; R^(3b)is selected from the group consisting of H and D; R⁴ is a memberselected from the group consisting of H and C₁₋₈ alkyl, wherein the C₁₋₈alkyl is optionally substituted with —CONR^(a)R^(b), —OC(O)NR^(a)R^(b),—NR^(a)C(O)R^(b), —NR^(a)C(O)₂R^(c), —NR^(a)R^(b), or —OR^(a), whereineach R^(a) and R^(b) is independently selected from hydrogen, C₁₋₄alkyl, C₁₋₄ hydroxyalkyl and C₁₋₄ haloalkyl, and R^(c) is selected fromC₁₋₄ alkyl, C₁₋₄ hydroxyalkyl and C₁₋₄ haloalkyl; R^(5a) and R^(5b) areeach members independently selected from the group consisting of H, F,Cl and CH₃; R^(6a) and R^(6b) are each members independently selectedfrom the group consisting of H, C₁₋₄ alkyl, C₁₋₄ hydroxyalkyl and C₁₋₄haloalkyl; or optionally R^(6a) and R^(6b) are taken together to formoxo (═O); or any salts, solvates, hydrates, N-oxides or rotamersthereof.

Within formula (Ib), further selected embodiments are those in whichR^(3b) is H; R⁴ is H or CH₃; R^(5a) is H, F or Cl; R^(5b) is H, F, Cl;R^(6a) and R^(6b) are independently selected from the group consistingof H and CH₃, or are taken together to form oxo (═O).

Within formula (Ib), still other selected embodiments are those in whichR^(3b) is D; R⁴ is H or CH₃; R^(5a) is H, F or Cl; R^(5b) is H, F, Cl;R^(6a) and R^(6b) are independently selected from the group consistingof H and CH₃, or are taken together to form oxo (═O).

In another selected group of embodiments, the compound is selected fromthose provided in the Examples below, or in Table 1.

In each of the selected embodiments, the noted compounds may be presentin a pharmaceutically acceptable salt or hydrate form.

In some embodiments, a compound or a pharmaceutically acceptable saltthereof is provided selected from the group consisting of:

Still further, for those compounds shown above without stereochemistry,the present invention is also directed to chiral forms of each of thecompounds, as well as enantiomerically enriched forms of the notedcompounds. Enantiomerically enriched forms can be prepared using chiralchromatography according to well-known methods practiced in the art or,for example, by chiral resolution with a chiral salt form. In someembodiments, the enantiomeric excess for an enantiomerically enrichedform is at least 10%, 20%, 30%, 40%, 50%, 60% or more. In still otherembodiments, an enantiomerically enriched form is provided that is atleast 70%, 80%, 90%, 95%, or more.

Preparation of Compounds

Certain compounds of the invention can be prepared following methodologyas described in the Examples section of this document. In addition thesyntheses of certain intermediate compounds that are useful in thepreparation of compounds of the invention are also described.

B. Compositions

In addition to the compounds provided above, compositions for modulatingCXCR2 activity in humans and animals will typically contain apharmaceutical carrier or diluent.

The term “composition” as used herein is intended to encompass a productcomprising the specified ingredients in the specified amounts, as wellas any product which results, directly or indirectly, from combinationof the specified ingredients in the specified amounts. By“pharmaceutically acceptable” it is meant the carrier, diluent orexcipient must be compatible with the other ingredients of theformulation and not deleterious to the recipient thereof.

The pharmaceutical compositions for the administration of the compoundsof this invention may conveniently be presented in unit dosage form andmay be prepared by any of the methods well known in the art of pharmacyand drug delivery. All methods include the step of bringing the activeingredient into association with the carrier which constitutes one ormore accessory ingredients. In general, the pharmaceutical compositionsare prepared by uniformly and intimately bringing the active ingredientinto association with a liquid carrier or a finely divided solid carrieror both, and then, if necessary, shaping the product into the desiredformulation. In the pharmaceutical composition the active objectcompound is included in an amount sufficient to produce the desiredeffect upon the process or condition of diseases.

The pharmaceutical compositions containing the active ingredient may bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsions and self-emulsifications as described in U.S. PatentApplication 2002-0012680, hard or soft capsules, syrups, elixirs,solutions, buccal patch, oral gel, chewing gum, chewable tablets,effervescent powder and effervescent tablets. Compositions intended fororal use may be prepared according to any method known to the art forthe manufacture of pharmaceutical compositions and such compositions maycontain one or more agents selected from the group consisting ofsweetening agents, flavoring agents, coloring agents, antioxidants andpreserving agents in order to provide pharmaceutically elegant andpalatable preparations. Tablets contain the active ingredient inadmixture with non-toxic pharmaceutically acceptable excipients whichare suitable for the manufacture of tablets. These excipients may be forexample, inert diluents, such as cellulose, silicon dioxide, aluminumoxide, calcium carbonate, sodium carbonate, glucose, mannitol, sorbitol,lactose, calcium phosphate or sodium phosphate; granulating anddisintegrating agents, for example, corn starch, or alginic acid;binding agents, for example PVP, cellulose, PEG, starch, gelatin oracacia, and lubricating agents, for example magnesium stearate, stearicacid or talc. The tablets may be uncoated or they may be coated,enterically or otherwise, by known techniques to delay disintegrationand absorption in the gastrointestinal tract and thereby provide asustained action over a longer period. For example, a time delaymaterial such as glyceryl monostearate or glyceryl distearate may beemployed. They may also be coated by the techniques described in theU.S. Pat. Nos. 4,256,108; 4,166,452; and U.S. Pat. No. 4,265,874 to formosmotic therapeutic tablets for control release.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin, or olive oil.Additionally, emulsions can be prepared with a non-water miscibleingredient such as oils and stabilized with surfactants such asmono-diglycerides, PEG esters and the like.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydroxy-propylmethyl cellulose, sodiumalginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxy-ethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl, p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, forexample olive oil or arachis oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative and flavoring and coloringagents. Oral solutions can be prepared in combination with, for example,cyclodextrin, PEG and surfactants.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally-acceptable diluent orsolvent, for example as a solution in 1,3-butane diol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose any bland fixed oil may be employedincluding synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid find use in the preparation of injectables.

The compounds provided herein may also be administered in the form ofsuppositories for rectal administration of the drug. These compositionscan be prepared by mixing the drug with a suitable non-irritatingexcipient which is solid at ordinary temperatures but liquid at therectal temperature and will therefore melt in the rectum to release thedrug. Such materials include cocoa butter and polyethylene glycols.Additionally, the compounds can be administered via ocular delivery bymeans of solutions or ointments. Still further, transdermal delivery ofthe subject compounds can be accomplished by means of iontophoreticpatches and the like. For topical use, creams, ointments, jellies,solutions or suspensions, etc., containing the compounds provided hereinare employed. As used herein, topical application is also meant toinclude the use of mouth washes and gargles.

The compounds of this invention may also be coupled a carrier that is asuitable polymers as targetable drug carriers. Such polymers can includepolyvinylpyrrolidone, pyran copolymer,polyhydroxy-propyl-methacrylamide-phenol,polyhydroxyethyl-aspartamide-phenol, or polyethyleneoxide-polylysinesubstituted with palmitoyl residues. Furthermore, the compounds of theinvention may be coupled to a carrier that is a class of biodegradablepolymers useful in achieving controlled release of a drug, for examplepolylactic acid, polyglycolic acid, copolymers of polylactic andpolyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid,polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates andcross linked or amphipathic block copolymers of hydrogels. Polymers andsemipermeable polymer matrices may be formed into shaped articles, suchas valves, stents, tubing, prostheses and the like.

A pharmaceutical composition comprising a compound of the presentdisclosure is provided. In some embodiments, the pharmaceuticalcomposition further comprises one or more additional therapeutic agents.In some embodiments, the one or more additional therapeutic agent isselected from the group consisting of a cytotoxic chemotherapy,anti-cancer or anti-tumor vaccines, anti-immunocytokine therapies,immunocytokine therapies, chimeric antigen receptor (CAR) T cellreceptors, gene transfer therapy, and checkpoint inhibitors. In someembodiments, the one or more additional therapeutic agent is selectedfrom the group consisting of: drugs that block the activity of CTLA-4(CD152), PD-1 (CD279), PDL-1 (CD274), TIM-3, LAG-3 (CD223), VISTA, KIR,NKG2A, BTLA, PD-1H, TIGIT, CD96, 4-1BB (CD137), 4-1BBL (CD137L), GARP,CSF-1R, A2AR, CD73, CD47, tryptophan 2,3-dioxygenase (TDO) orindoleamine 2,3 dioxygenase (IDO), and agonists of OX40, GITR, 4-1BB,ICOS, STING or CD40.

C. Methods of Use

While not wishing to be bound by any particular theory, the compoundsand compositions provided herein are considered to provide a therapeuticeffect by inhibiting the CXCR2 receptor. Therefore, the compounds andcompositions provided herein can be used in the treatment or preventionof diseases or disorders in a mammal in which the inhibition of theCXCR2 receptor would provide a therapeutic effect.

Another aspect of the invention is the use of a compound as providedherein and/or the pharmaceutically acceptable salt and/or a prodrugthereof alone or in combination with other medicaments or activeingredients as for producing a medicament for the treatment orprophylaxis of a chemokine mediated disease, wherein the chemokine bindsto a CXCR2 receptor.

Accordingly, provided herein are methods directed to the use of acompound of formula I and/or a pharmaceutically acceptable salt and/or aprodrug thereof alone or in combination with other medicaments or activeingredients as for producing a medicament for the treatment orprophylaxis of rheumatoid arthritis, chronic obstructive pulmonarydisease, adult or acute respiratory distress syndrome, asthma,atherosclerosis, myocardial and renal ischemia/reperfusion injury,peripheral limb ischemia/reperfusion injury, inflammatory bowel disease,ulcerative colitis, Crohn's disease, meconium aspriration syndrome,atopic dermatitis, cystic fibrosis, psoriasis, psoriatic arthritis,multiple sclerosis, angiogenesis, restenosis, osteoarthritis,osteoporosis, septic shock, endotoxic shock, gram negative sepsis, toxicshock syndrome, stroke, glomerulonephritis, thrombosis, graft vs. hostreaction, allograft rejections, transplant reperfusion injury, earlytransplantation rejection, acute inflammation, Alzheimer's disease,malaria, respiratory viruses, herpes viruses, hepatitis viruses, HIV,Kaposi's sarcoma-associated viruses, meningitis, gingivitis, herpesencephalitis, CNS vasculitis, traumatic brain injury, brainischemia/reperfusion injury, migraine, CNS tumors, subarachnoidhemorrhage, post-surgical trauma, interstitial pneumonitis,hypersensitivity, crystal induced arthritis, acute and chronicpancreatitis, hepatic ischemia/reperfusion injury, acute alcoholichepatitis, necrotizing enterocolitis, chronic sinusitis, uveitis,polymyositis, vasculitis, acne, gastric and duodenal ulcers, intestinalischemia/reperfusion injury, celiac disease, esophagitis, glossitis,rhinitis, airflow obstruction, airway hyper-responsiveness,bronchiolitis, bronchiolitis obliterans, bronchiolitis obliteransorganizing pneumonia, bronchiectasis, chronic bronchitis, cor pulmonae,dyspnea, emphysema, hypercapnea, hyperinflation, hyperoxia-inducedinflammations, hypoxemia, hypoxia, lung ischemia/reperfusion injury,pulmonary fibrosis, pulmonary hypertension, right ventricularhypertrophy, peritonitis associated with continuous ambulatoryperitoneal dialysis, granulocytic ehrlichiosis, sarcoidosis, smallairway disease, ventilation-perfusion mismatching, wheeze, colds, gout,alcoholic liver disease, lupus, burn therapy, periodontitis, pre-termlabor, cough, pruritis, multi-organ dysfunction, trauma, sprains,contusions, undesired hematopoietic stem cell release, angiogenic oculardisease, ocular inflammation, retinopathy or prematurity, diabeticretinopathy, macular degeneration with the wet type preferred andcorneal neovascularization, tumor angiogenesis, cancer and metastasis.

In particular, the invention further relates to the use of a compound ofthe formula I and/or a pharmaceutically acceptable salt and/or a prodrugthereof alone or in combination with other medicaments or activeingredients as for producing a medicament for the treatment orprophylaxis of acute and chronic inflammatory diseases such asatherosclerosis, ischemia/reperfusion injuries, chronic obstructivepulmonary disease, asthma, and rheumatoid arthritis, chemokine (such as,but not limited to IL-8, GRO-α, GRO-β, GRO-γ, NAP-2, ENA-78, or GCP-2)mediated diseases which include adult respiratory distress syndrome,inflammatory bowel disease, ulcerative colitis, Crohn's disease, atopicdermatitis, cystic fibrosis, psoriasis, dermatitis, multiple sclerosis,angiogenesis, restenosis, osteoarthritis, septic shock, endotoxic shock,gram negative sepsis, toxic shock syndrome, stroke, glomerulonephritis,thrombosis, graft vs. host reaction, allograft rejections, Alzheimer'sdisease, malaria, viral infections, traumatic brain injury, pulmonaryfibrosis, and cancer.

In some embodiments, the compounds and compositions of the invention areadministered to a subject having cancer. In some cases, CXCR2 inhibitorsare administered to treat cancer, e.g., carcinomas, gliomas,mesotheliomas, melanomas, lymphomas, leukemias (including acutelymphocytic leukemias), adenocarcinomas, breast cancer, ovarian cancer,cervical cancer, glioblastoma, leukemia, lymphoma, prostate cancer, andBurkitt's lymphoma, head and neck cancer, colon cancer, colorectalcancer, non-small cell lung cancer, small cell lung cancer, cancer ofthe esophagus, stomach cancer, pancreatic cancer, hepatobiliary cancer,cancer of the gallbladder, cancer of the small intestine, rectal cancer,kidney cancer, renal cancer, bladder cancer, prostate cancer, penilecancer, urethral cancer, testicular cancer, cervical cancer, vaginalcancer, uterine cancer, ovarian cancer, thyroid cancer, parathyroidcancer, adrenal cancer, pancreatic endocrine cancer, carcinoid cancer,bone cancer, skin cancer, retinoblastomas, Hodgkin's lymphoma,non-Hodgkin's lymphoma (see, CANCER: PRINCIPLES AND PRACTICE (DeVita, V.T. et al. eds 1997) for additional cancers); as well as brain andneuronal dysfunction, such as Alzheimer's disease, multiple sclerosisand demyelinating diseases; hypertensive disorders such as pulmonaryarterial hypertension; kidney dysfunction; renal dysfunction; rheumatoidarthritis; allograft rejection; atherosclerosis (and elevatedcholesterol levels); asthma; glomerulonephritis; contact dermatitis;inflammatory bowel disease; colitis; psoriasis; reperfusion injury; aswell as other disorders and diseases described herein. In someembodiments, the subject does not have Kaposi's sarcoma, multicentricCastleman's disease or AIDS-associated primary effusion lymphoma.

In some embodiments, a method of treating a CXCR2-mediated disease orcondition in a subject in need thereof is provided, said methodcomprising administering an effective amount of a compound of thepresent disclosure, or a pharmaceutically acceptable salt thereof, or apharmaceutical composition of the present disclosure to said subject. Insome embodiments, the CXCR2-mediated disease is an acute or chronicinflammatory disorder. In some embodiments, the CXCR2-mediated acute orchronic inflammatory disorder is selected from the group consisting ofpsoriasis, rheumatoid arthritis, radiation induced fibrotic lungdisease, autoimmune bullous dermatosis (AIBD), chronic obstructivepulmonary disease, and ozone-induced airway inflammation.

In some embodiments, a compound of the present disclosure, or apharmaceutically acceptable salt thereof, is used to treat cancer aloneor in combination with one or more other anti-cancer therapies. In someembodiments, a compound of the present disclosure or a pharmaceuticallyacceptable salt thereof, is used to treat cancer in combination with oneor more of a cytotoxic chemotherapy, an anti-cancer vaccine, ananti-tumor vaccines, an anti-immunocytokine, an immunocytokine therapy,and a chimeric antigen receptor (CAR) T cell receptors, gene transfertherapy. In some embodiments, a compound of the present disclosure or apharmaceutically acceptable salt thereof, is used to treat cancer incombination with one or more checkpoint inhibitor. In some embodiments,a compound of the present disclosure is used to treat cancer incombination with one or more of an anti-cancer therapy selected from thegroup consisting of drugs that block the activity of CTLA-4 (CD152),PD-1 (CD279), PDL-1 (CD274), TIM-3, LAG-3 (CD223), VISTA, KIR, NKG2A,BTLA, PD-1H, TIGIT, CD96, 4-1BB (CD137), 4-1BBL (CD137L), GARP, CSF-1R,A2AR, CD73, CD47, tryptophan 2,3-dioxygenase (TDO) or indoleamine 2,3dioxygenase (IDO), and agonists of OX40, GITR, 4-1BB, ICOS, STING orCD40.

In some embodiments, the compounds of the disclosure, or apharmaceutically acceptable salt and/or a prodrug thereof, orcompositions of the disclosure are administered to treat melanoma,glioblastoma, esophagus tumor, nasopharyngeal carcinoma, uveal melanoma,lymphoma, lymphocytic lymphoma, primary CNS lymphoma, T-cell lymphoma,diffuse large B-cell lymphoma, primary mediastinal large B-celllymphoma, prostate cancer, castration-resistant prostate cancer, chronicmyelocytic leukemia, Kaposi's sarcoma fibrosarcoma, liposarcoma,chondrosarcoma, osteogenic sarcoma, angiosarcoma, lymphangiosarcoma,synovioma, meningioma, leiomyosarcoma, rhabdomyosarcoma, sarcoma of softtissue, sarcoma, sepsis, biliary tumor, basal cell carcinoma, thymusneoplasm, cancer of the thyroid gland, cancer of the parathyroid gland,uterine cancer, cancer of the adrenal gland, liver infection, Merkelcell carcinoma, nerve tumor, follicle center lymphoma, colon cancer,Hodgkin's disease, non-Hodgkin's lymphoma, leukemia, chronic or acuteleukemias including acute myeloid leukemia, chronic myeloid leukemia,acute lymphoblastic leukemia, chronic lymphocytic leukemia, multiplemyeloma, ovary tumor, myelodysplastic syndrome, cutaneous or intraocularmalignant melanoma, renal cell carcinoma, small-cell lung cancer, lungcancer, mesothelioma, breast cancer, squamous non-small cell lung cancer(SCLC), non-squamous NSCLC, colorectal cancer, ovarian cancer, gastriccancer, hepatocellular carcinoma, pancreatic carcinoma, pancreaticcancer, Pancreatic ductal adenocarcinoma, squamous cell carcinoma of thehead and neck, cancer of the head or neck, gastrointestinal tract,stomach cancer, bone cancer, skin cancer, rectal cancer, cancer of theanal region, testicular cancer, carcinoma of the fallopian tubes,carcinoma of the endometrium, carcinoma of the cervix, carcinoma of thevagina, carcinoma of the vulva, cancer of the esophagus, cancer of thesmall intestine, cancer of the endocrine system, cancer of the urethra,cancer of the penis, cancer of the bladder, cancer of the kidney, cancerof the ureter, carcinoma of the renal pelvis, neoplasm of the centralnervous system (CNS), tumor angiogenesis, spinal axis tumor, brain stemglioma, pituitary adenoma, epidermoid cancer, abestosis, carcinoma,adenocarcinoma, papillary carcinoma, cystadenocarcinoma, bronchogeniccarcinoma, renal cell carcinoma, transitional cell carcinoma,choriocarcinoma, seminoma, embryonal carcinoma, wilm's tumor,pleomorphic adenoma, liver cell papilloma, renal tubular adenoma,cystadenoma, papilloma, adenoma, leiomyoma, rhabdomyoma, hemangioma,lymphangioma, osteoma, chondroma, lipoma and/or fibroma.

Other disorders involving unwanted or problematic angiogenesis includerheumatoid arthritis; psoriasis; ocular angiogenic diseases, forexample, diabetic retinopathy, retinopathy of prematurity, maculardegeneration, corneal graft rejection, neovascular glaucoma, retrolentalfibroplasia, rubeosis; Osler-Webber Syndrome; myocardial angiogenesis;plaque neovascularization; telangiectasia; hemophiliac joints;angiofibroma; disease of excessive or abnormal stimulation ofendothelial cells, including intestinal adhesions, Crohn's disease, skindiseases such as psoriasis, eczema, and scleroderma, diabetes, diabeticretinopathy, retinopathy of prematurity, age-related maculardegeneration, atherosclerosis, scleroderma, wound granulation andhypertrophic scars, i.e., keloids, and diseases that have angiogenesisas a pathologic consequence such as cat scratch disease and ulcers(Helicobacter pylori), can also be treated with antibodies of theinvention. Angiogenic inhibitors can be used to prevent or inhibitadhesions, especially intra-peritoneal or pelvic adhesions such as thoseresulting after open or laparoscopic surgery, and bum contractions.Other conditions which should be beneficially treated using theangiogenesis inhibitors include prevention of scarring followingtransplantation, cirrhosis of the liver, pulmonary fibrosis followingacute respiratory distress syndrome or other pulmonary fibrosis of thenewborn, implantation of temporary prosthetics, and adhesions aftersurgery between the brain and the dura. Endometriosis, polyposis,cardiac hypertrophy, as well as obesity, may also be treated byinhibition of angiogenesis. These disorders may involve increases insize or growth of other types of normal tissue, such as uterinefibroids, prostatic hypertrophy, and amyloidosis. Compounds andcompositions provided herein may be used prophylactically ortherapeutically for any of the disorders or diseases described herein.

In some embodiments, the compounds of the disclosure, or apharmaceutically acceptable salt and/or a prodrug thereof, orcompositions of the disclosure are administered to treat cystitis,insulin dependent diabetes, islet cell transplant rejection; kidneytransplant rejection; liver transplant rejection; lung transplantrejection, COPD, or influenza.

Methods of Treating Cancer

More specifically, the present invention also provides a method oftreating cancer. A preferred method of treating cancer, includesadministering a therapeutically effective amount of one or more of thepreviously mentioned compounds (or salts thereof) to a cancer patientfor a time sufficient to treat the cancer.

For treatment, the compositions provided herein may be administered byoral, parenteral (e.g., intramuscular, intraperitoneal, intravenous,ICV, intracisternal injection or infusion, subcutaneous injection, orimplant), by inhalation spray, nasal, vaginal, rectal, sublingual, ortopical routes of administration and may be formulated, alone ortogether, in suitable dosage unit formulations containing conventionalnon-toxic pharmaceutically acceptable carriers, adjuvants and vehiclesappropriate for each route of administration.

In some embodiments, selective CXCR2 inhibitors provided herein can beadministered in combination with other appropriate therapeutic agents,including, e.g., chemotherapeutic agents, radiation, etc. It isunderstood that such administration may be prior to, subsequent to or inunison with the second therapeutic agent, such that the therapeuticeffects of the second agent are enhanced when compared to administrationof the second agent in the absence of the CXCR2 inhibitor. Selection ofthe appropriate agents for use in combination therapy may be made by oneof ordinary skill in the art, according to conventional pharmaceuticalprinciples. The combination of therapeutic agents may actsynergistically to effect the treatment or prevention of the variousdisorders such as, e.g., cancer, wounds, kidney dysfunction, braindysfunction or neuronal dysfunction. Using this approach, one may beable to achieve therapeutic efficacy with lower dosages of each agent,thus reducing the potential for adverse side effects.

In addition to primates, such as humans, a variety of other mammals canbe treated according to the method provided herein. For instance,mammals including, but not limited to, cows, sheep, goats, horses, dogs,cats, guinea pigs, rats or other bovine, ovine, equine, canine, feline,rodent or murine species can be treated. However, the method can also bepracticed in other species, such as avian species (e.g., chickens).

Standard in vivo assays demonstrating that the compositions providedherein are useful for treating cancer include those described inBertolini, F., et al., Endostatin, an antiangiogenic drug, induces tumorstabilization after chemotherapy or anti-CD20 therapy in a NOD/SCIDmouse model of human high-grade non-Hodgkin lymphoma. Blood, No. 1, Vol.96, pp. 282-87 (1 Jul. 2000); Pengnian, L., Antiangiogenic gene therapytargeting the endothelium-specific receptor tyrosine kinase Tie2. Proc.Natl. Acad. Sci. USA, Vol. 95, pp. 8829-34 (July 1998); and Pulaski, B.Cooperativity of Staphylococcal aureus Enterotoxin B Superantigen, MajorHistocompatibility Complex Class II, and CD80 for ImmunotherapyofAdvanced Spontaneous Metastases in a Clinically Relevant PostoperativeMouse Breast Cancer Model. Cancer Research, Vol. 60, pp. 2710-15 (May15, 2000).

In the treatment or prevention of conditions which require chemokinereceptor modulation an appropriate dosage level will generally be about0.001 to 100 mg per kg patient body weight per day which can beadministered in single or multiple doses. Preferably, the dosage levelwill be about 0.01 to about 25 mg/kg per day; more preferably about 0.05to about 10 mg/kg per day. A suitable dosage level may be about 0.01 to25 mg/kg per day, about 0.05 to 10 mg/kg per day, or about 0.1 to 5mg/kg per day. Within this range the dosage may be 0.005 to 0.05, 0.05to 0.5 or 0.5 to 5.0 mg/kg per day. For oral administration, thecompositions are preferably provided in the form of tablets containing1.0 to 1000 milligrams of the active ingredient, particularly 1.0, 5.0,10.0, 15.0. 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0,400.0, 500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0 milligrams of theactive ingredient for the symptomatic adjustment of the dosage to thepatient to be treated. The compounds may be administered on a regimen of1 to 4 times per day, preferably once or twice per day.

It will be understood, however, that the specific dose level andfrequency of dosage for any particular patient may be varied and willdepend upon a variety of factors including the activity of the specificcompound employed, the metabolic stability and length of action of thatcompound, the age, body weight, hereditary characteristics, generalhealth, sex and diet of the subject, as well as the mode and time ofadministration, rate of excretion, drug combination, and the severity ofthe particular condition for the subject undergoing therapy.

The compounds and compositions provided herein can be combined withother compounds and compositions having related utilities to prevent andtreat cancer and diseases or conditions associated with CXCR2 signaling.Such other drugs may be administered, by a route and in an amountcommonly used therefor, contemporaneously or sequentially with acompound or composition provided herein. When a compound or compositionprovided herein is used contemporaneously with one or more other drugs,a pharmaceutical composition containing such other drugs in addition tothe compound or composition provided herein is preferred. Accordingly,the pharmaceutical compositions provided herein include those that alsocontain one or more other active ingredients or therapeutic agents, inaddition to a compound or composition provided herein. Examples of othertherapeutic agents that may be combined with a compound or compositionprovided herein, either administered separately or in the samepharmaceutical compositions, include, but are not limited to: cisplatin,paclitaxel, methotrexate, cyclophosphamide, ifosfamide, chlorambucil,carmustine, carboplatin, vincristine, vinblastine, thiotepa, lomustine,semustine, 5-fluorouracil and cytarabine. The weight ratio of thecompound provided herein to the second active ingredient may be variedand will depend upon the effective dose of each ingredient. Generally,an effective dose of each will be used. Thus, for example, when acompound provided herein is combined with a second anticancer agent, theweight ratio of the compound provided herein to the second agent willgenerally range from about 1000:1 to about 1:1000, preferably about200:1 to about 1:200. Combinations of a compound provided herein andother active ingredients will generally also be within theaforementioned range, but in each case, an effective dose of each activeingredient should be used. In some embodiments, the compounds of thedisclosure, or a pharmaceutically acceptable salt and/or a prodrugthereof, are administered either separately or in the samepharmaceutical compositions, with: an alkylation agent, a nitrosoureaagent, an anticancer antibiotics, a vegetable-origin alkaloid, atopoisomerase inhibitor, an hormone drug, an hormone antagonist, anaromatase inhibitor, a P-glycoprotein inhibitor, a platinum complexderivative, an immunotherapeutic drug or another anticancer drugs, orany combination thereof.

Methods of Treating Inflammation

Still further, the compounds and compositions provided herein are usefulfor the treatment of inflammation, and can be combined with othercompounds and compositions having therapeutic utilities that may requiretreatment either before, after or simultaneously with the treatment ofcancer or inflammation with the present compounds. Accordingly,combination methods and compositions are also a component of the presentinvention to prevent and treat the condition or disease of interest,such as inflammatory or autoimmune disorders, conditions and diseases,including inflammatory bowel disease, rheumatoid arthritis,osteoarthritis, psoriatic arthritis, polyarticular arthritis, multiplesclerosis, allergic diseases, psoriasis, atopic dermatitis and asthma,and those pathologies noted above.

For example, in the treatment or prevention of inflammation orautoimmunity or for example arthritis associated bone loss, the presentcompounds and compositions may be used in conjunction with ananti-inflammatory or analgesic agent such as an opiate agonist, alipoxygenase inhibitor, such as an inhibitor of 5-lipoxygenase, acyclooxygenase inhibitor, such as a cyclooxygenase-2 inhibitor, aninterleukin inhibitor, such as an interleukin-1 inhibitor, an NMDAantagonist, an inhibitor of nitric oxide or an inhibitor of thesynthesis of nitric oxide, a non-steroidal anti-inflammatory agent, or acytokine-suppressing anti-inflammatory agent, for example with acompound such as acetaminophen, aspirin, codeine, fentanyl, ibuprofen,indomethacin, ketorolac, morphine, naproxen, phenacetin, piroxicam, asteroidal analgesic, sufentanyl, sunlindac, tenidap, and the like.Similarly, the instant compounds and compositions may be administeredwith an analgesic listed above; a potentiator such as caffeine, an H2antagonist (e.g., ranitidine), simethicone, aluminum or magnesiumhydroxide; a decongestant such as phenylephrine, phenylpropanolamine,pseudoephedrine, oxymetazoline, epinephrine, naphazoline,xylometazoline, propylhexedrine, or levo desoxy ephedrine; anantitussive such as codeine, hydrocodone, caramiphen, carbetapentane, ordextromethorphan; a diuretic; and a sedating or non-sedatingantihistamine.

As noted, compounds and compositions provided herein may be used incombination with other drugs that are used in the treatment, prevention,suppression or amelioration of the diseases or conditions for whichcompounds and compositions provided herein are useful. Such other drugsmay be administered, by a route and in an amount commonly used therefor,contemporaneously or sequentially with a compound or composition asprovided herein. When a compound or composition as provided herein isused contemporaneously with one or more other drugs, a pharmaceuticalcomposition containing such other drugs in addition to the compound orcomposition provided herein is preferred. Accordingly, thepharmaceutical compositions provided herein include those that alsocontain one or more other active ingredients or therapeutic agents, inaddition to a compound or composition as provided herein. Examples ofother therapeutic agents that may be combined with a compound orcomposition provided herein, either administered separately or in thesame pharmaceutical compositions, include, but are not limited to: (a)VLA-4 antagonists, (b) corticosteroids, such as beclomethasone,methylprednisolone, betamethasone, prednisone, prenisolone,dexamethasone, fluticasone, hydrocortisone, budesonide, triamcinolone,salmeterol, salmeterol, salbutamol, formeterol; (c) immunosuppressantssuch as cyclosporine (cyclosporine A, Sandimmune®, Neoral®), tacrolirnus(FK-506, Prograf®), rapamycin (sirolimus, Rapamune®) and other FK-506type immunosuppressants, and rnycophenolate, e.g., mycophenolate mofetil(CellCept®); (d) antihistamines (H1-histamine antagonists) such asbromopheniramine, chlorpheniramine, dexchloipheniramine, triprolidine,clemastine, diphenhydramine, diphenylpyraline, tripelennamine,hydroxyzine, methdilazine, promethazine, trimeprazine, azatadine,cyproheptadine, antazoline, pheniramine pyrilamine, astemizole,terfenadine, loratadine, cetirizine, fexofenadine,descarboethoxyloratadine, and the like; (e) non steroidal antiasthmatics (e.g., terbutaline, metaproterenol, fenoterol, isoetharine,albuterol, bitolterol and pirbuterol), theophylline, cromolyn sodium,atropine, ipratropium bromide, leukotriene antagonists (e.g.,zafmlukast, montelukast, pranlukast, iralukast, pobilukast andSKB-106,203), leukotriene biosynthesis inhibitors (zileuton, BAY-1005);(f) non-steroidal anti-inflammatory agents (NSAIDs) such as propionicacid derivatives (e.g., alminoprofen, benoxaprofen, bucloxic acid,carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen, ibuprofen,indoprofen, ketoprofen, rniroprofen, naproxen, oxaprozin, pirprofen,pranoprofen, suprofen, tiaprofenic acid and tioxaprofen), acetic acidderivatives (e.g., indomethacin, acemetacin, alclofenac, clidanac,diclofenac, fenclofenac, fenclozic acid, fentiazac, furofenac, ibufenac,isoxepac, oxpinac, sulindac, tiopinac, tolmetin, zidometacin andzomepirac), fenamic acid derivatives (e.g., flufenamic acid,meclofenamic acid, mefenamic acid, niflumic acid and tolfenamic acid),biphenylcarboxylic acid derivatives (e.g., diflunisal and flufenisal),oxicams (e.g., isoxicam, piroxicam, sudoxicam and tenoxican),salicylates (e.g., acetyl salicylic acid and sulfasalazine) and thepyrazolones (e.g., apazone, bezpiperylon, feprazone, mofebutazone,oxyphenbutazone and phenylbutazone); (g) cyclooxygenase-2 (COX-2)inhibitors such as celecoxib (Celebrex®) and rofecoxib (Vioxx®); (h)inhibitors of phosphodiesterase type IV (PDE IV); (i) gold compoundssuch as auranofin and aurothioglucose, (j) etanercept (Enbrel®), (k)antibody therapies such as orthoclone (OKT3), daclizumab (Zenapax®),basiliximab (Simulect®) and infliximab (Remicade®), (l) otherantagonists of the chemokine receptors, especially CCR5, CXCR2, CXCR3,CCR2, CCR3, CCR4, CCR7, CX₃CR1 and CXCR6; (m) lubricants or emollientssuch as petrolatum and lanolin, (n) keratolytic agents (e.g.,tazarotene), (o) vitamin D₃ derivatives, e.g., calcipotriene orcalcipotriol (Dovonex®), (p) PUVA, (q) anthralin (Drithrocreme®), (r)etretinate (Tegison®) and isotretinoin and (s) multiple sclerosistherapeutic agents such as interferon β-1β (Betaseron®), interferon(β-1α (Avonex®), azathioprine (Imurek®, Imuran®), glatiramer acetate(Capoxone®), a glucocorticoid (e.g., prednisolone) and cyclophosphamide(t) DMARDS such as methotrexate (u) other compounds such as5-aminosalicylic acid and prodrugs thereof; hydroxychloroquine;D-penicillamine; antimetabolites such as azathioprine, 6-mercaptopurineand methotrexate; DNA synthesis inhibitors such as hydroxyurea andmicrotubule disrupters such as colchicine. The weight ratio of thecompound provided herein to the second active ingredient may be variedand will depend upon the effective dose of each ingredient. Generally,an effective dose of each will be used. Thus, for example, when acompound provided herein is combined with an NSAID the weight ratio ofthe compound provided herein to the NSAID will generally range fromabout 1000:1 to about 1:1000, preferably about 200:1 to about 1:200.Combinations of a compound provided herein and other active ingredientswill generally also be within the aforementioned range, but in eachcase, an effective dose of each active ingredient should be used.

Method of Diagnosing Diseases and Disorders Associated with CXCR2

Still further, the compounds and compositions provided herein are usefulfor the diagnosis of diseases and disorders associated with CXCR2. Inparticular, the compounds provided herein can be prepared in a labeledform (e.g., radiolabeled) and used for the diagnosis of, for example,cancer. Labeled compounds provided herein that bind to CXCR2 (e.g.,antagonists or agonists) can be used to determine levels of CXCR2 in amammalian subject. In some embodiments, the CXCR2 modulators orantagonists are administered to a subject having cancer. In some cases,labeled compounds are administered to detect developing cancers, e.g.,carcinomas, gliomas, mesotheliomas, melanomas, lymphomas, leukemias,adenocarcinomas, breast cancer, ovarian cancer, cervical cancer,glioblastoma, leukemia, lymphoma, prostate cancer, and Burkitt'slymphoma, head and neck cancer, colon cancer, colorectal cancer,non-small cell lung cancer, small cell lung cancer, cancer of theesophagus, stomach cancer, pancreatic cancer, hepatobiliary cancer,cancer of the gallbladder, cancer of the small intestine, rectal cancer,kidney cancer, bladder cancer, prostate cancer, penile cancer, urethralcancer, testicular cancer, cervical cancer, vaginal cancer, uterinecancer, ovarian cancer, thyroid cancer, parathyroid cancer, adrenalcancer, pancreatic endocrine cancer, carcinoid cancer, bone cancer, skincancer, retinoblastomas, Hodgkin's lymphoma, non-Hodgkin's lymphoma(see, CANCER: PRINCIPLES AND PRACTICE (DeVita, V. T. et al. eds 1997)for additional cancers); as well as brain and neuronal dysfunction, suchas Alzheimer's disease and multiple sclerosis; kidney dysfunction;rheumatoid arthritis; cardiac allograft rejection; atherosclerosis (andelevated cholesterol levels); asthma; glomerulonephritis; contactdermatitis; inflammatory bowel disease; colitis; psoriasis; reperfusioninjury; as well as other disorders and diseases described herein. Insome embodiments, the subject does not have Kaposi's sarcoma,multicentric Castleman's disease or AIDS-associated primary effusionlymphoma.

A variety of imaging and detection methods can be used for the detectionof cancers. In some embodiments, direct methods are available toevaluate CXCR2 biodistribution in the body such as magnetic resonanceimaging (“MRI”), positron emission tomography (“PET”), and single photonemission computed tomography (“SPECT”). Each of these methods can detectthe distribution of a suitably labeled compound (generally as bound toCXCR2) within the body if that compound contains an atom with theappropriate nuclear properties. MRI detects paramagnetic nuclei; PET andSPECT detect the emission of particles from the decay of radionuclei.

For methods involving PET, it is necessary to incorporate an appropriatepositron-emitting radionuclide. There are relatively fewpositron-emitting isotopes that are suitable for labeling a therapeuticagent. The carbon isotope, ¹¹C, has been used for PET, but has a shorthalf-life of 20.5 minutes. Accordingly, the facilities for synthesis anduse are typically near to a cyclotron where the precursor ¹¹C startingmaterial is generated. Another useful isotope, ¹⁸F, has a half-life of110 minutes. This allows sufficient time for incorporation into aradiolabeled tracer, for purification and for administration into ahuman or animal subject. Other isotopes have even shorter half-lives.¹³N has a half-life of 10 minutes and ¹⁵O has an even shorter half-lifeof 2 minutes. The emissions of both are more energetic, however, thanthose of ¹¹C and PET studies have been carried out with these isotopes(see, Clinical Positron Emission Tomography, Mosby Year Book, 1992, K.F. Hubner, et al., Chapter 2).

SPECT imaging employs isotope tracers that are 7-emitters. While therange of useful isotopes is greater than for PET, imaging with SPECTprovides lower three-dimensional resolution. However, in some instances,SPECT is used to obtain clinically significant information aboutcompound binding, localization and clearance rates. One useful isotopefor SPECT imaging is ¹²³I, a 7-emitter with a 13.3 hour half-life.Compounds labeled with ¹²³I can be shipped up to about 1000 miles fromthe manufacturing site, or the isotope itself can be transported foron-site synthesis. Eighty-five percent of the isotope's emissions are159 KeV photons, which are readily measured by SPECT instrumentationcurrently in use. Other halogen isotopes can serve for PET or SPECTimaging, or for conventional tracer labeling. These include ⁷⁵Br, ⁷⁶Br,⁷⁷Br and ⁸²Br as having usable half-lives and emission characteristics.

In view of the above, the present invention provides methods for imaginga tumor, organ, or tissue, said method comprising:

-   -   (a) administering to a subject in need of such imaging, a        radiolabeled or detectable form of a compound of Formula I; and    -   (b) detecting said compound to determine where said compound is        concentrated in said subject.

Additionally, the present invention provides methods for detectingelevated levels of CXCR2 in a sample, said method comprising:

-   -   (a) contacting a sample suspected of having elevated levels of        CXCR2 with a radiolabeled or detectable form of a compound of        Formula I;    -   (b) determining a level of compound that is bound to CXCR2        present in said sample to determine the level of CXCR2 present        in said sample; and    -   (c) comparing the level determined in step (b) with a control        sample to determine if elevated levels of CXCR2 are present in        said sample.

As with the treatment methods described herein, administration of thelabeled compounds can be by any of the routes normally used forintroducing a compound into ultimate contact with the tissue to beevaluated and is well known to those of skill in the art. Although morethan one route can be used to administer a particular composition, aparticular route can often provide a more immediate and more effectivediagnosis than another route.

Combination Therapies

Inhibitors of CXCR2 can be supplied alone or in conjunction with one ormore other drugs. Possible combination partners can include, e.g.,additional anti-angiogenic factors and/or chemotherapeutic agents (e.g.,cytotoxic agents) or radiation, a cancer vaccine, an immunomodulatoryagent, a checkpoint inhibitor, an anti-vascular agent, a signaltransduction inhibitor, an antiproliferative agent, or an apoptosisinducer.

Examples of other therapeutic agents that may be combined with acompound or composition of the present disclosure, either administeredseparately or in the same pharmaceutical composition, include, but arenot limited to: modulators of CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7,CCR8, CCR9, CCR10, CCR11, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6,CXCR7, CX3CR1, ChemR23, C5aR, C5a, and C5, or any combination thereof.In some embodiments, the modulator is an antagonist.

Examples of other therapeutic agents that may be combined with acompound or composition of the present disclosure, either administeredseparately or in the same pharmaceutical compositions, include, but arenot limited to: a therapeutic antibody, a bispecific antibody and“antibody-like” therapeutic protein (such as DARTs®, Duobodies®, Bites®,XmAbs®, TandAbs®, Fab derivatives), an antibody-drug conjugate (ADC), avirus, an oncolytic virus, gene modifiers or editors such as CRISPR(including CRISPR Cas9), zinc finger nucleases or synthetic nucleases(TALENs), a CAR (chimeric antigen receptor) T-cell immunotherapeuticagent, cytokines, vaccines, vaccine adjuvants, GM-CSF, M-CSF, G-CSF,interferon-a, beta, or gamma, IL-1, IL-2, IL-3, IL-12, Poly (I:C), CPG,cyclophosphamide, analogs of cyclophosphamide, anti-TGF and imatinib(Gleevac), a mitosis inhibitor paclitaxel, Sunitinib (Sutent),antiangiogenic agents, an aromatase inhibitor, letrozole, an A2aadenosine receptor (A2AR) antagonist, an adenosine receptor modulator,an A3 adenosine receptor modulator, an angiogenesis inhibitor,anthracyclines, oxaliplatin, doxorubicin, TLR4 antagonists, IL-18antagonists, a Btk tyrosine kinase inhibitor, an Erbb2 tyrosine kinasereceptor inhibitor; an Erbb4 tyrosine kinase receptor inhibitor, an mTORinhibitor, a Thymidylate synthase inhibitor, an EGFR tyrosine kinasereceptor inhibitor, an Epidermal growth factor antagonist, a Fyntyrosine kinase inhibitor, a Kit tyrosine kinase inhibitor, a Lyntyrosine kinase inhibitor, a NK cell receptor modulator, a PDGF receptorantagonist, a PARP inhibitor, a Poly ADP ribose polymerase inhibitor, aPoly ADP ribose polymerase 1 inhibitor, a Poly ADP ribose polymerase 2inhibitor, a Poly ADP ribose polymerase 3 inhibitor, aGalactosyltransferase modulator, a Dihydropyrimidine dehydrogenaseinhibitor, an Orotate phosphoribosyltransferase inhibitor, a Telomerasemodulator, a Mucin 1 inhibitor, a Mucin inhibitor, a Secretin agonist, aTNF related apoptosis inducing ligand modulator, an IL17 genestimulator, an Interleukin 17E ligand, a Neurokinin receptor agonist, aCyclin G1 inhibitor, a checkpoint inhibitor, a PD-1 inhibitor, a PD-L1inhibitor, a CTLA4 inhibitor, a Topoisomerase I inhibitor, an Alk-5protein kinase inhibitor, a Connective tissue growth factor ligandinhibitor, a Notch-2 receptor antagonist, a Notch-3 receptor antagonist,a Hyaluronidase stimulator, a MEK-1 protein kinase inhibitor, aPhosphoinositide-3 kinase inhibitor, a MEK-2 protein kinase inhibitor, aGM-CSF receptor modulator; TNF alpha ligand modulator, a Mesothelinmodulator, an Asparaginase stimulator, a CSF2 gene stimulator, aCaspase-3 stimulator; Caspase-9 stimulator, a PKN3 gene inhibitor, aHedgehog protein inhibitor; Smoothened receptor antagonist, an AKT1 geneinhibitor, a DHFR inhibitor, a Thymidine kinase stimulator, a CD29modulator, a Fibronectin modulator, an Interleukin-2 ligand, a Serineprotease inhibitor, a D40LG gene stimulator; TNFSF9 gene stimulator, a 2oxoglutarate dehydrogenase inhibitor, a TGF-beta type II receptorantagonist, an Erbb3 tyrosine kinase receptor inhibitor, aCholecystokinin CCK2 receptor antagonist, a Wilms tumor proteinmodulator, a Ras GTPase modulator, an Histone deacetylase inhibitor, aRaf B protein kinase inhibitor, a Cyclin-dependent kinase 4 inhibitor Amodulator, an Estrogen receptor beta modulator, a 4-1BB inhibitor, a4-1BBL inhibitor, a PD-L2 inhibitor, a B7-H3 inhibitor, a B7-H4inhibitor, a BTLA inhibitor, a HVEM inhibitor, a TIM3 inhibitor, a TIGITinhibitor, a NKG2A inhibitor, a GAL9 inhibitor, a LAG3 inhibitor, aPD-1H inhibitor, a PD96 inhibitor, a VISTA inhibitor, a KIR inhibitor, a2B4 inhibitor, a CD160 inhibitor, a CD66e modulator, an Angiotensin IIreceptor antagonist, a Connective tissue growth factor ligand inhibitor,a Jak1 tyrosine kinase inhibitor, a Jak2 tyrosine kinase inhibitor, adual Jak1/Jak2 tyrosine kinase inhibitor, an Angiotensin convertingenzyme 2 stimulator, a Growth hormone receptor antagonist, a Galectin-3inhibitor, a Checkpoint kinase 2 modulator, a Sodium glucosetransporter-2 inhibitor, a Endothelin ET-A antagonist, aMineralocorticoid receptor antagonist, an Endothelin ET-B antagonist, anAdvanced glycosylation product receptor antagonist, anAdrenocorticotrophic hormone ligand, a Farnesoid X receptor agonist, aG-protein coupled bile acid receptor 1 agonist, an Aldose reductaseinhibitor, a Xanthine oxidase inhibitor, a PPAR gamma agonist, aProstanoid receptor antagonist, a FGF receptor antagonist, a PDGFreceptor antagonist, a TGF beta antagonist, a P3 protein modulator, ap38 MAP kinase inhibitor, a VEGF-1 receptor antagonist, a Proteintyrosine phosphatase beta inhibitor, a Tek tyrosine kinase receptorstimulator, a PDE 5 inhibitor, a Mineralocorticoid receptor antagonist,an ACE inhibitor, a I-kappa B kinase inhibitor, a NFE2L2 genestimulator, a Nuclear factor kappa B inhibitor, a STAT3 gene inhibitor,a NADPH oxidase 1 inhibitor, a NADPH oxidase 4 inhibitor, a PDE 4inhibitor, a Renin inhibitor, a FURIN gene inhibitor, a MEKK-5 proteinkinase inhibitor, a Membrane copper amine oxidase inhibitor, an Integrinalpha-V/beta-3 antagonist, an Insulin sensitizer, a Kallikrein 1modulator, a Cyclooxygenase inhibitor, a Complement C3 modulator, aTubulin binding agent, a Macrophage mannose receptor 1 modulator, aPhenylalanine hydroxylase stimulator, an OX40 agonist, a GITR agonist, aCD40 agonist, Denileukin diftitox, Bexarotene, Vorinostat, Romidepsin,Pralatrexate, prednisone, prednisolone, CCX354, CCX9588, CCX140, CCX872,CCX598, CCX6239, CCX9664, CCX2553, CCX 2991, CCX282, CCX025, CCX507,CCX430, CCX765, CCX224, CCX662, CCX650, CCX832, CCX168, CCX168-M1,bavituximab, IMM-101, CAP1-6D, Rexin-G, genistein, CVac, MM-D37K,PCI-27483, TG-01, mocetinostat, LOAd-703, CPI-613, upamostat, CRS-207,NovaCaps, trametinib, Atu-027, sonidegib, GRASPA, trabedersen,nastorazepide, Vaccell, oregovomab, istiratumab, refametinib,regorafenib, lapatinib, selumetinib, rucaparib, pelareorep, tarextumab,PEGylated hyaluronidase, varlitinib, aglatimagene besadenovec, GBS-01,GI-4000, WF-10, galunisertib, afatinib, RX-0201, FG-3019, pertuzumab,DCVax-Direct, selinexor, glufosfamide, virulizin, yttrium (90Y)clivatuzumab tetraxetan, brivudine, nimotuzumab, algenpantucel-L,tegafur+gimeracil+oteracil potassium+calcium folinate, olaparib,ibrutinib, pirarubicin, Rh-Apo2L, tertomotide,tegafur+gimeracil+oteracil potassium, tegafur+gimeracil+oteracilpotassium, masitinib, Rexin-G, mitomycin, erlotinib, adriamycin,dexamethasone, vincristine, cyclophosphamide, fluorouracil, topotecan,taxol, interferons, platinum derivatives, taxane, paclitaxel, vincaalkaloids, vinblastine, anthracyclines, doxorubicin,epipodophyllotoxins, etoposide, cisplatin, rapamycin, methotrexate,actinomycin D, dolastatin 10, colchicine, emetine, trimetrexate,metoprine, cyclosporine, daunorubicin, teniposide, amphotericin,alkylating agents, chlorambucil, 5-fluorouracil, campthothecin,cisplatin, metronidazole, Gleevec, Avastin, Vectibix, abarelix,aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine,amifostine, anastrozole, arsenic trioxide, asparaginase, azacitidine,AZD9291, BCG Live, bevacuzimab, fluorouracil, bexarotene, bleomycin,bortezomib, busulfan, calusterone, capecitabine, camptothecin,carboplatin, carmustine, celecoxib, cetuximab, chlorambucil, cladribine,clofarabine, cyclophosphamide, cytarabine, dactinomycin, darbepoetinalfa, daunorubicin, denileukin, dexrazoxane, docetaxel, doxorubicin(neutral), doxorubicin hydrochloride, dromostanolone propionate,epirubicin, epoetin alfa, estramustine, etoposide phosphate, etoposide,exemestane, filgrastim, floxuridine fludarabine, fulvestrant, gefitinib,gemcitabine, gemtuzumab, goserelin acetate, histrelin acetate,hydroxyurea, ibritumomab, idarubicin, ifosfamide, imatinib mesylate,interferon alfa-2a, interferon alfa-2b, irinotecan, lenalidomide,letrozole, leucovorin, leuprolide acetate, levamisole, lomustine,megestrol acetate, melphalan, 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propanediol, pegvisomant, GR-MD-02,canagliflozin, irbesartan, FG-3019, atrasentan, finerenone, sparsentan,bosentan, defibrotide, fimasartan, azeliragon, pyridoxamine,corticotropin, INT-767, epalrestat, topiroxostat, SER-150-DN,pirfenidone, VEGFR-1 mAb, AKB-9778, PF-489791, SHP-627, CS-3150,imidapril, perindopril, captopril, enalapril, lisinopril, Zofenopril,Lisinopril, Quinapril, Benazepril, Trandolapril, Cilazapril, Fosinopril,Ramipril, bardoxolone methyl, irbesartan+propagermanium, GKT-831,MT-3995, TAK-648, TAK-272, GS-4997, DW-1029M, ASP-8232, VPI-2690B,DM-199, rhein, PHN-033, GLY-230, and sapropterin, sulodexide, lirilumab,IPH-4102, IPH-2101, IMP-321, BMS-986016, MGD-013, LAG-525, durvalumab,monalizumab, MCLA-134, MBG-453, CA-170, AUPM-170, AUPM-327, resminostat,ipilimumab, BGB-A317, tremelimumab, REGN-2810, AZD-5069, masitinib,binimetinib, trametinib, ruxolitinib, dabrafenib, linaclotide,ipilimumab, apatinib, nintedanib, cabozantinib, pazopanib, belinostat,panitumumab, guadecitabine, vismodegib, vemurafenib, dasatinib,tremelimumab, bevacizumab, oxaliplatin, aflibercept, vandetanib,everolimus, thalidomide, veliparib, encorafenib, napabucasin, alpelisib,axitinib, cediranib, necitumumab, ramucirumab, irofulven,trifluridine+tipiracil, donafenib, pacritinib, pexastimogenedevacirepvec, tivantinib, GNR-011, talaporfin, piclidenoson, decitabine,ganitumab, panobinostat, rintatolimod, polmacoxib, levofolinate,famitinib, votumumab, tivozanib, entinostat, plitidepsin, lefitolimod,OSE-2101, vitespen, TroVax, bromocriptine, midostaurin, fosbretabulin,fruquintinib, ganetespib, brivanib, anlotinib, L19-TNF-alpha,racotumomab, Novaferon, raltitrexed, enzastaurin, GM-CT-01, arcitumomab,or any combination thereof.

Kits and Packages

The terms “kit” and “pharmaceutical kit” refer to a commercial kit orpackage comprising, in one or more suitable containers, one or morepharmaceutical compositions and instructions for their use. In oneembodiment, kits comprising a compound of Formula (I), or apharmaceutically acceptable salt thereof, and instructions for itsadministration are provided. In one embodiment, kits comprising acompound of Formula (I), or a pharmaceutically acceptable salt thereof,in combination with one or more (e.g., one, two, three, one or two, orone to three) additional therapeutic agents and instructions for theiradministration are provided.

In one embodiment, the compounds of this disclosure are formulated intoadministration units which are packaged in a single packaging. Thesingle packaging encompasses but is not limited to a bottle, achild-resistant bottle, an ampoule, and a tube. In one embodiment, thecompounds of this disclosure and optionally additional therapeuticagents, are formulated into administration units and every singleadministration unit is individually packaged in a single packaging. Suchindividually packaged units may contain the pharmaceutical compositionin any form including but not limited to liquid form, solid form, powderform, granulate form, an effervescent powder or tablet, hard or softcapsules, emulsions, suspensions, syrup, suppositories, tablet, troches,lozenges, solution, buccal patch, thin film, oral gel, chewable tablet,chewing gum, and single-use syringes. Such individually packaged unitsmay be combined in a package made of one or more of paper, cardboard,paperboard, metal foil and plastic foil, for example a blister pack. Oneor more administration units may be administered once or several times aday. One or more administration units may be administered three times aday. One or more administration units may be administered twice a day.One or more administration units may be administered on a first day andone or more administration units may be administered on the followingdays.

General Synthetic Procedure

The embodiments are also directed to processes and intermediates usefulfor preparing the subject compounds or pharmaceutically acceptable saltsthereof.

Exemplary chemical entities useful in methods of the embodiments willnow be described by reference to illustrative synthetic schemes fortheir general preparation herein and the specific examples that follow.Artisans will recognize that, to obtain the various compounds herein,starting materials may be suitably selected so that the ultimatelydesired substituents will be carried through the reaction scheme with orwithout protection as appropriate to yield the desired product.Alternatively, it may be necessary or desirable to employ, in the placeof the ultimately desired substituent, a suitable group that may becarried through the reaction scheme and replaced as appropriate with thedesired substituent. Furthermore, one of skill in the art will recognizethat the transformations shown in the schemes below may be performed inany order that is compatible with the functionality of the particularpendant groups.

Representative syntheses of compounds of the present disclosure aredescribed in the schemes below, and the particular examples that follow.Schemes 1 and 2 are provided as further embodiment of the disclosure andillustrate general methods which were used to prepare compounds of thepresent disclosure including compounds of Formula (I), (Ia) and (Ib),and which can be used to prepare additional compounds having the Formula(I), (Ia), and (Ib). The methodology is compatible with a wide varietyof functionalities.

The amino group of A1 can be reacted with3,4-dimethoxycyclobut-3-ene-1,2-dione to provide A2. A2 can then bereacted with the amino group of A3 to provide A4.

A7 can be obtained by reduction of the cyano group in A6, for example byhydrogenation, followed by cyclization. Alternatively, AS (where Xrepresents a leaving group such as a halogen or a tosylate and where Ris an alkyl group), can be reacted with NH₃ to form the cyclized productA7. A7 can be reacted with HNO₃ to introduce the nitro group in presenceof an acid such as sulfuric acid to give A8. Subsequent reduction of thenitro group in A8 by for example hydrogenation can provide A9.

EXAMPLES

The following examples are offered to illustrate, but not to limit theclaimed invention.

Reagents and solvents used below can be obtained from commercial sourcessuch as Aldrich Chemical Co. (Milwaukee, Wis., USA). ¹H-NMR spectra wererecorded on a Varian Mercury 400 MHz NMR spectrometer. Significant peaksare provided relative to TMS and are tabulated in the order:multiplicity (s, singlet; d, doublet; t, triplet; q, quartet; m,multiplet) and number of protons. Mass spectrometry results are reportedas the ratio of mass over charge, followed by the relative abundance ofeach ion (in parenthesis). In the examples, a single m/e value isreported for the M+H (or, as noted, M−H) ion containing the most commonatomic isotopes. Isotope patterns correspond to the expected formula inall cases. Electrospray ionization (ESI) mass spectrometry analysis wasconducted on a Hewlett-Packard MSD electrospray mass spectrometer usingthe HP 1100 HPLC for sample delivery. Normally the analyte was dissolvedin methanol at 0.1 mg/mL and 1 microlitre was infused with the deliverysolvent into the mass spectrometer, which scanned from 100 to 1500daltons. All compounds could be analyzed in the positive ESI mode, usingacetonitrile/water with 1% formic acid as the delivery solvent. Thecompounds provided below could also be analyzed in the negative ESImode, using 2 mM NH₄OAc in acetonitrile/water as delivery system.

The following abbreviations are used in the Examples and throughout thedescription of the invention: rt, room temperature; HPLC, high pressureliquid chromatography; TFA, trifluoroacetic acid; LC-MSD, liquidchromatograph/mass selective detector; LC-MS, liquid chromatograph/massspectrometer; Pd₂dba₃, tris(dibenzylideneacetone) dipalladium; THF,tetrahydrofuran; DMF, dimethylformamide or N,N-dimethylformamide; DCM,dichloromethane; DMSO, dimethyl sulfoxide; TLC, thin-layerchromatography; KHMDS, potassium hexamethyldisilazane; ES, electrospray;sat., saturated.

Compounds within the scope of this invention can be synthesized asdescribed below, using a variety of reactions known to the skilledartisan. One skilled in the art will also recognize that alternativemethods may be employed to synthesize the target compounds of thisinvention, and that the approaches described within the body of thisdocument are not exhaustive, but do provide broadly applicable andpractical routes to compounds of interest.

Certain molecules claimed in this patent can exist in differentenantiomeric and diastereomeric forms and all such variants of thesecompounds are claimed.

The detailed description of the experimental procedures used tosynthesize key compounds in this text lead to molecules that aredescribed by the physical data identifying them as well as by thestructural depictions associated with them.

Those skilled in the art will also recognize that during standard workup procedures in organic chemistry, acids and bases are frequently used.Salts of the parent compounds are sometimes produced, if they possessthe necessary intrinsic acidity or basicity, during the experimentalprocedures described within this patent.

Example 1: Synthesis of3-[(5-fluoro-3-oxo-isoindolin-4-yl)amino]-4-[[(1R)-1-(5-methyl-2-furyl)propyl]amino]cyclobut-3-ene-1,2-dione

Step a:

A 500 mL round-bottom flask was charged with methyl2-bromo-5-fluorobenzoate (48 g, 206 mmol), copper cyanide (37 g, 412mmmol) and DMF (200 mL). The mixture was heated at 110° C. overnight andthen cooled to room temperature. Ether (1.5 L) and Celite (100 g) wereadded and the mixture was stirred at room temperature for 30 minutes.The solid was filtered and the filtrate was washed with brine (3×200 mL)and then dried over MgSO₄. The solvent was evaporated under reducedpressure to give the desired product as a colorless solid (31 g, 84%).MS: (ES) m/z calculated for C₉H₇FNO₂[M+H]⁺ 180, found 180.

Step b:

To a solution of methyl 2-cyano-5-fluorobenzoate (10 g, 56 mmol) inmethanol (200 mL) was added 10% Pd—C (1.0 g) at room temperature. Theresulting mixture was stirred under a hydrogen (50 psi) atmosphereovernight. The reaction mixture was filtered through Celite and thefiltrate was concentrated under reduced pressure to give the desiredproduct as a colorless solid (8.0 g, 90%). MS: (ES) m/z calculated forC₈H₇FNO[M+H]⁺ 152, found 152.

Step c:

To a 0° C. suspension of 6-fluoroisoindoline-1-one (8.0 g, 5.3 mmol) inconcentrated H₂SO₄ was added drop-wise a pre-cooled mixture ofconcentrated H₂SO₄ (26 mL) and nitric acid (6 mL) while keeping thereaction mixture below 5° C. After addition, the reaction mixture wasslowly warmed to room temperature during overnight. Ice (50 g) was addedto the mixture and the solid was collected and dried, then washed withMTBE (50 mL) and ethyl acetate (50 mL) to give the desired product as alight yellow solid (5.1 g, 50%). MS: (ES) m/z calculated forC₈H₆FN₂O₃[M+H]⁺ 197, found 197.

Step d:

A solution of 6-fluoro-7-nitroisoindoline-1-one (11.3 g, 57 mmol) and10% Pd/C (50% wet, 6.2 g, 2.9 mmol, 0.05 equiv) in THF (300 mL) wasstirred under a hydrogen atmosphere (balloon) overnight. The solid wasfiltered through Celite and the filtrate was concentrated under reducedpressure to give a colorless solid, which was purified by silica gelchromatography (100% ethyl acetate) to give the desired product as awhite solid (6.4 g, 67%). MS: (ES) m/z calculated for C₈H₉FN₂O[M+H]⁺168, found 168.

Step e:

A mixture of 7-amino-6-difluoro-isoindolin-1-one (4.4 g, 26 mmol) and3,4-dimethoxycyclobut-3-ene-1,2-dione (7.4 g, 52 mmol) in anhydrousmethanol (30 mL) was stirred at 60° C. for overnight and then at 80° C.for 5 h. The reaction mixture was evaporated and the residue was stirredin ethyl acetate (200 mL) at 50° C. for 30 min, then cooled down to roomtemperature. The mixture was filtered and dried to give a light yellowcolor solid (5.0 g, 70%). MS: (ES) m/z calculated for C₁₃H₁₀FN₂O₄ [M+H]⁺277, found 277.

Step f:

A 20 mL vial was charged with3-[(5-fluoro-3-oxo-isoindolin-4-yl)amino]-4-methoxy-cyclobut-3-ene-1,2-dione(29.8 mg, 0.108 mmol), followed by(1R)-1-(5-methyl-2-furyl)propan-1-amine (25.2 mg, 0.181 mmol) in ethanol(1 mL). The reaction mixture was stirred at ambient temperatureovernight. After gently blowing nitrogen over the reaction mixture toremove most of the solvent, dichloromethane and 1N-hydrochloric acidwere added to the reaction mixture. The organic layer was separated andthe aqueous layer was extracted twice more with dichloromethane. Thecombined organic layers were dried over anhydrous sodium sulfate. Afterremoval of the solvent under reduced pressure, the crude material waspurified using silica gel column chromatography using a mixture ofdichloromethane and ethyl acetate as the eluent.3-[(5-Fluoro-3-oxo-isoindolin-4-yl)amino]-4-[[(1R)-1-(5-methyl-2-furyl)propyl]amino]cyclobut-3-ene-1,2-dione(27.8 mg, 0.0725 mmol) was obtained in 67% yield. ¹H NMR (400 MHz,DMSO-d₆) δ 9.49 (s, 1H), 8.72 (s, 1H), 8.26 (d, J=9.1 Hz, 1H), 7.47 (dd,J=11.2, 8.3 Hz, 1H), 7.34 (dd, J=8.3, 3.8 Hz, 1H), 6.26 (d, J=3.1 Hz,1H), 6.05 (dd, J=3.0, 1.3 Hz, 1H), 5.08 (dd, J=8.1, 8.1 Hz, 1H), 4.33(s, 2H), 2.26 (s, 3H), 1.91 (ddq, J=28, 8.1, 7.3 Hz, 2H), 0.91 (t, J=7.3Hz, 3H). MS: (ES) m/z calculated for C₂₀H₁₈FN₃O₄ [M+H]⁺ 384.1, found384.3.

Example 2: Synthesis of3-[(5-fluoro-3-oxo-isoindolin-4-yl)amino]-4-[[(1R)-2-methyl-1-(5-methyl-2-furyl)propyl]amino]cyclobut-3-ene-1,2-dione

Step a:

To a 500 mL round bottom flask was added(S)-2-methylpropane-2-sulfinamide (12.1 g, 100 mmol) followed bydichloromethane (100 mL) at ambient temperature.5-Methylfuran-2-carboxaldehyde (10.9 mL, 110 mmol) in dichloromethane(13 mL) and titanium ethoxide (51 mL, 219 mmol) in dichloromethane (87mL) were added. The reaction mixture was stirred overnight. The reactionwas diluted with dichloromethane (150 mL) and quenched with sodiumsulfate decahydrate (51 g). The reaction mixture was filtered throughcelite and rinsed with dichloromethane. Evaporation of the solvent gavethe crude(S)-2-methyl-N-[(5-methyl-2-furyl)methylene]propane-2-sulfinamide (20.89g, 97.9 mmol) which was used directly in the next reaction.

Step b:

(S)-2-methyl-N-[(5-methyl-2-furyl)methylene]propane-2-sulfinamide (1.76g, 8.19 mmol) was dissolved in toluene (40 mL) and the reaction wascooled to −70° C. using a dry ice/isopropyl alcohol bath. Isopropylmagnesium chloride (8.2 mL, 2M solution in THF, 16.4 mmol) was addedover 10 minutes. The reaction was gradually warmed to ambienttemperature and stirred overnight. The reaction was quenched by theaddition of saturated aqueous ammonium chloride. Organic materials wereextracted using diethyl ether three times followed by a brine wash. Theorganic layer was dried over anhydrous sodium sulfate and after removalof the solvent under reduced pressure, the crude mixture was purifiedusing a silica gel column using methyl tert-butyl ether/dichloromethaneas eluent.(S)-2-Methyl-N-[(1R)-2-methyl-1-(5-methyl-2-furyl)propyl]propane-2-sulfinamidewas obtained after removal of solvent under reduced pressure (530 mg2.06 mmol, 90% de from NMR).

Step c:

Acetyl chloride (0.366 mL, 5.15 mmol) was added dropwise to methanol (5mL) at 0° C. to prepare a solution of anhydrous hydrogen chloride inmethanol. This solution was added to(S)-2-methyl-N-[(1R)-2-methyl-1-(5-methyl-2-furyl)propyl]propane-2-sulfinamide(530 mg, 2.06 mmol) at 0° C. The reaction was slowly warmed to ambienttemperature over 2 hours. Saturated sodium bicarbonate solution wasadded to neutralize the reaction mixture and the product was extractedwith dichloromethane four times. The combined organic layer was driedover anhydrous sodium sulfate. Removal of the solvent under reducedpressure afforded the crude material (310 mg, 2.03 mmol) which was useddirectly in the next reaction.

Step d:

To3-[(5-fluoro-3-oxo-isoindolin-4-yl)amino]-4-methoxy-cyclobut-3-ene-1,2-dione(42.3 mg, 0.146 mmol) was added(1R)-2-methyl-1-(5-methyl-2-furyl)propan-1-amine (30.5 mg, 0.199 mmol)in ethanol (1 mL) at ambient temperature. The reaction mixture wasstirred overnight at 45° C. followed by stirring at 65° C. for 5 hours.A stream of nitrogen was gently blown over the reaction mixture toremove most of the solvent. Dichloromethane and 1N-hydrochloric acid wasadded and the layers were separated. The aqueous layer was extractedtwice more with dichloromethane. The combined organic layers were driedover anhydrous sodium sulfate. Solvent was removed under reducedpressure and the crude material was purified by silica gel columnchromatography using a mixture of dichloromethane and ethyl acetate asthe eluent.3-[(5-Fluoro-3-oxo-isoindolin-4-yl)amino]-4-[[(1R)-2-methyl-1l-(5-methyl-2-furyl)propyl]amino]cyclobut-3-ene-1,2-dione(52.4 mg, 0.132 mmol) was obtained in 90% yield. ¹H NMR (400 MHz,DMSO-d₆) δ 9.55 (s, 1H), 8.71 (s, 1H), 8.31 (d, J=9.7 Hz, 1H), 7.48 (dd,J=11.1, 8.3 Hz, 1H), 7.34 (dd, J=8.3, 3.8 Hz, 1H), 6.22 (d, J=3.1 Hz,1H), 6.05 (d, J=3.0 Hz, 1H), 4.97 (t, J=8.5 Hz, 1H), 4.33 (s, 2H), 2.27(s, 3H), 2.18 (dt, J=13.7, 6.8 Hz, 1H), 0.96 (d, J=6.7 Hz, 3H), 0.88 (d,J=6.7 Hz, 3H). MS: (ES) m/z calculated for C₂₁H₂₀FN₃O₄ [M+H]⁺398.2,found 398.4.

Example 3: Synthesis of3-[(5,7-difluoro-3-oxo-isoindolin-4-yl)amino]-4-[[(1R)-1-(5-methyl-2-furyl)propyl]amino]cyclobut-3-ene-1,2-dione

Step a:

3,5-Difluoro-2-methyl-benzoic acid (5.2 g, 30.2 mmol) was dissolved inanhydrous DMF (30 mL). Anhydrous Na₂CO₃ (3.5 g, 33.2 mmol, 1.1 equiv)was added and the reaction was stirred at room temperature for 30minutes. Methyl iodide (2.1 mL, 33.2 mmol, 1.1 equiv) was added and themixture was stirred at room temperature for 4 h, then the reaction wasdiluted with water (200 mL) and the product was extracted using Et₂O(3×50 mL). The combined organic layers were washed with brine (4×30 mL),dried over MgSO₄, filtered and evaporated to give a yellow oil (5.4 g,96%).

Step b:

The product from Step a (5.4 g, 29.0 mmol) was dissolved in carbontetrachloride (60 mL) and N-bromosuccinimide (7.7 g, 43.5 mmol, 1.5equiv) was added followed by benzoyl peroxide (1.4 g, 5.8 mmol, 0.20equiv). The reaction mixture was stirred at reflux overnight then cooledto room temperature and filtered. The filtrate was evaporated and theresidue was purified by column chromatography (silica gel, 100% hexanesto 9:1 hexanes: ethyl acetate) to give the product as a yellow oil (7.4g, 96%).

Step c:

NH₃ in methanol (7 M, 45 mL, 6.4 mmol) was cooled to 0° C. and theproduct from Step b (6 g, 22.6 mmol) was added. The reaction mixture wasstirred at 0° C. for 10 minutes and then at room temperature overnight.Excess solvent was evaporated and the residue was diluted with water (50mL). The resulting solid was filtered and washed with water (2×20 mL),then hexanes (20 mL) to give the product (3.4 g, 89%). MS: (ES) m/zcalculated for C₈H₆F₂NO [M+H]⁺ 170.04, found 170.3.

Step d:

The 4,6-difluoroisoindolin-1-one from Step c (3.4 g, 20.1 mmol) wasdissolved in concentrated H₂SO₄ (40 mL) and cooled to 0° C. 70% HNO₃(1.5 mL, 24.1 mmol, 1.2 equiv) was added drop-wise and the reactionmixture was stirred at 0° C. for 10 minutes, then allowed to warm toroom temperature over a period of 1 hour and stirred overnight. Ice wasadded and the mixture was then diluted with cold water (100 mL). Theresulting yellow solid was filtered, washed with water (2×50 mL), thenhexanes (50 mL) and dried under vacuum (3.4 g, 79%). MS: (ES) m/zcalculated for C₈H₅F₂N₂O₃ [M+H]⁺ 215.03, found 215.2.

Step e:

The 4,6-difluoro-7-nitro-isoindolin-1-one from Step d (3.4 g, 15.9 mmol)was diluted with THF (50 mL) and 10% Pd/C, 50% wet, (1.7 g, 0.8 mmol, 5%mmol) was added under a nitrogen atmosphere. The reaction mixture wasvigorously stirred under H₂ (balloon) for 1 day at room temperature,then filtered through Celite and evaporated to give the solid product(2.7 g, 92%). MS: (ES) m/z calculated for C₈H₇F₂N₂O [M+H]⁺ 185.05, found185.3.

Step f:

A mixture of 7-amino-4,6-difluoro-isoindolin-1-one from Step e (2.3 g,12.5 mmol) and 3,4-dimethoxycyclobut-3-ene-1,2-dione (3.5 g, 25.0 mmol,2.0 equiv) in anhydrous MeOH (15 mL) was stirred at 60° C. overnight.The reaction mixture was evaporated and the residue was diluted withMTBE:EtOAc (1:1, 200 mL) and stirred at 50° C. for 30 min, then cooleddown to room temperature. The solid product was filtered, washed withMTBE, then dissolved in MeOH:DCM (1:1, 200 mL) and filtered throughCelite. The filtrate was evaporated to give a gray solid (2.0 g, 54%).MS: (ES) m/z calculated for C₁₃H₉F₂N₂O₄ [M+H]⁺ 295.05, found 295.3.

Step g:

A 20 mL vial was charged with3-[(5,7-difluoro-3-oxo-isoindolin-4-yl)amino]-4-methoxy-cyclobut-3-ene-1,2-dione(44.3 mg, 0.144 mmol), followed by(1R)-1-(5-methyl-2-furyl)propan-1-amine (25 mg, 0.18 mmol) in ethanol (1mL). The reaction mixture was stirred at ambient temperature overnight,then at 80° C. for 2 hours. After gently blowing nitrogen over thereaction mixture to remove most of the solvent, dichloromethane and1N-hydrochloric acid were added to the reaction mixture. The organiclayer was separated and the aqueous layer was extracted twice more withdichloromethane. The combined organic layers were dried over anhydroussodium sulfate. After removal of the solvent under reduced pressure, thecrude material was purified by silica gel column chromatography using amixture of dichloromethane and ethyl acetate as the eluent. The obtainedproduct was re-suspended in ethyl acetate and filtered. The solid wasfiltered and dried under vacuum to give3-[(5,7-Difluoro-3-oxo-isoindolin-4-yl)amino]-4-[[(JR)-1-(5-methyl-2-furyl)propyl]amino]cyclobut-3-ene-1,2-dione(22.6 mg, 0.0563 mmol) in 39% yield. ¹H NMR (400 MHz, DMSO-d₆) δ 9.44(s, 1H), 8.91 (s, 1H), 8.19 (d, J=9.2 Hz, 1H), 7.62 (dd, J=10.8, 8.6 Hz,1H), 6.26 (d, J=3.0 Hz, 1H), 6.05 (d, J=3.0 Hz, 1H), 5.07 (dd, J=7.6,7.6 Hz, 1H), 4.41 (s, 2H), 2.26 (s, 3H), 1.90 (ddq, J=27, 7.6, 7.3 Hz,2H), 0.91 (t, J=7.3 Hz, 3H). MS: (ES) m/z calculated forC₂₀H₁₇F₂N₃O₄[M+H]⁺ 402.1, found 402.4.

Example 4: Synthesis of3-(((S)-5-fluoro-1-methyl-3-oxoisoindolin-4-yl)amino)-4-(((R)-1-(5-methylfuran-2-yl)propyl)amino)cyclobut-3-ene-1,2-dioneand3-(((R)-5-fluoro-1-methyl-3-oxoisoindolin-4-yl)amino)-4-(((R)-1-(5-methylfuran-2-yl)propyl)amino)cyclobut-3-ene-1,2-dione

Step a:

To a stirred solution of 6-fluoroisoindolin-1-one (10 g, 66.2 mmol) inanhydrous dichloromethane (100 mL) were added triethylamine (16.72,165.5 mmol, 21.8 mL), (Boc)₂O (17.3 g, 79.4 mmol) and catalytic DMAP(100 mg) at room temperature. The reaction mixture was stirred at roomtemperature for 16 h. After completion, the reaction mixture was dilutedwith CH₂Cl₂, washed with H₂O, and then saturated aqueous NaHCO₃. Theorganic layer was dried over Na₂SO₄, filtered, and concentrated invacuo. The crude compound was purified by silica gel chromatography(0-30% ethyl acetate in hexanes) to give the product. MS: (ES) m/zcalculated for C₁₃H₁₄FNO₃ [M+H]⁺ 252.3, found 252.3.

Step b:

1) To a stirred solution oftert-butyl-6-fluoro-1-oxoisoindoline-2-carboxylate (5.0 g, 19.9 mmol) inanhydrous THF (40 mL) at −78° C. under N₂ atmosphere was added LiHMDS(21.89 mL, 21.89 mmol) dropwise. After the solution was stirred for 30min, a solution of methyl idodide (2.82 g, 19.92 mmol) in THF (5 mL) wasadded to the mixture. The reaction mixture was stirred at −78° C. for 1h, and then the mixture was allowed to warm to room temperature andstirred for 2 h. After completion, the reaction mixture was quenchedwith saturated aqueous NH₄Cl, diluted with EtOAc (100 mL), and theorganic layer was washed with H₂O and then brine solution. The organiclayer was then dried over Na₂SO₄, filtered and concentrated in vacuo.The crude was used directly in the next step without any furtherpurification. 2) To a stirred solution oftert-butyl-5-fluoro-1-methyl-3-oxoisoindoline-2-carboxylate (6.2 g, 66.2mmol) in MeOH (60 mL) was added 4N HCl in dioxane (79.6 mmol, 20 mL).The mixture was stirred at room temperature for 3 h. After completion ofthe reaction, the solvent was removed and the reaction mixture wasdiluted with EtOAc (3×50 mL), the organic layer was washed with H₂O, andthen saturated aqueous NaHCO₃. The organic layer was then dried overNa₂SO₄, filtered and concentrated. The crude compound was purified bysilica gel, chromatography (10-80% ethyl acetate in hexanes) to give theproduct. MS: (ES) m/z calculated for C₉H₈FNO [M+H]⁺ 166.2, found 166.2.

Step c:

1) To a stirred solution of 6-fluoro-3-methylisoindolin-1-one (2.5 g,15.1 mmol) in anhydrous THF (25 mL) at −78° C. under N₂ atmosphere wasadded n-BuLi (6.64 mL, 16.61 mmol, 2.5 M in hexane) dropwise and thereaction mixture was stirred at −78° C. for 30 min, a solution of(1R,2S,5R)-2-isopropyl-5-methylcyclohexyl chloroformate (3.96 g, 18.18mmol) in THF (5 mL) was added to the mixture. The reaction mixture wasstirred at −78° C. for 30 min, then, the reaction mixture was allowed towarm to room temperature and stirred for 3 h. After completion of thereaction, the reaction mixture was quenched with satd. NH₄Cl, solution,extracted with EtOAc (2×75 mL), the combined organic layer was washedwith H₂O, and brine solution, dried over Na₂SO₄, filtered andconcentrated in vacuo. The crude compound was purified by silica gelchromatography to give (1S)-(1R,2S,5R)-2-isopropyl-5-methylcyclohexyl5-fluoro-1-methyl-3-oxoisoindoline-2-carboxylate and(1R)-(1R,2S,5R)-2-isopropyl-5-methylcyclohexyl5-fluoro-1-methyl-3-oxoisoindoline-2-carboxylate separately. 2) To astirred solution of one diastereomer obtained above (1.2 g, 3.45 mmol)in MeOH (10 mL) was added Mg(OMe)₂ (10-12% wt) in MeOH (17.2 mmol, 10mL) at room temperature. The reaction mixture was stirred at roomtemperature for 2 h. After completion of the reaction, the solvent wasremoved and the reaction mixture was quenched with satd. NH₄Cl,solution, extracted with EtOAc (2×75 mL), the combined organic layer waswashed with H₂O, and brine solution, dried over Na₂SO₄, filtered andconcentrated in vacuo. The crude compound was purified by silica gel,chromatography (20-60%) ethyl acetate/hexane to give the desiredproduct. MS: (ES) m/z calculated for C₉H₈FNO [M+H]⁺ 166.2, found 166.2.The other diastereomer was treated similarly to give the other desiredproduct.

Step d:

1) One of the compounds obtained from Step c (0.45 g, 2.72 mmol) wasdissolved in concentrated H₂SO₄ (5 mL) and cooled to 0° C. 70% HNO₃(0.34 g, 24.1 mmol, 2.0 equiv) was added drop-wise and the reactionmixture was stirred at 0° C. for 10 minutes, then allowed to warm toroom temperature stirred for overnight. Ice was added and the mixturewas then diluted with cold water (10 mL), the reaction mixture wasextracted with EtOAc (2×25 mL) washed with H₂O, and brine solution,dried over Na₂SO₄, filtered and concentrated. The crude was useddirectly in the next step without any further purification. MS: (ES) m/zcalculated for C₉H₇F₂N₂O₃ [M+H]⁺ 211.0, found 211.2. The otherenantiomer was treated similarly to give the other desired product. 2)One of the compounds obtained above (0.35 g, 1.32 mmol) and 10% Pd/C(50% wet, 100 mg) in MeOH (25 mL) was stirred under a hydrogenatmosphere (par shaker) for 1 h at 40 psi. The mixture was filteredthrough Celite and washed with MeOH (40 mL), the filtrate wasconcentrated under reduced pressure to give a colorless solid, which waspurified by silica gel chromatography (20-100% ethyl acetate/hexanes) togive the desired product. MS: (ES) m/z calculated for C₉H₉FN₂O[M+H]⁺181.1, found 181.2. The other enantiomer was treated similarly to givethe other desired product.

Step e:

A mixture of one of the compounds obtained in Step d (170 mg, 0.939mmol) and 3,4-dimethoxycyclobut-3-ene-1,2-dione (200 mg, 1.40 mmol) inanhydrous methanol (4 mL) was stirred at 60° C. for 3 h. The reactionmixture was evaporated and the residue was stirred in ethyl acetate (10mL) at 50° C. for 30 min, then cooled down to room temperature. Themixture was filtered and dried to give the desired product. MS: (ES) m/zcalculated for C₁₋₄H₁₁FN₂O₄ [M+H]⁺ 291.1, found 291.2. The otherenantiomer was treated similarly to give the other desired product.

Step f:

A 20 mL vial was charged with one of the compounds obtained above (29.0mg, 0.100 mmol), followed by (1R)-1-(5-methyl-2-furyl)propan-1-aminetartrate salt (28.9 mg, 0.100 mmol) in methanol (0.5 mL) andtriethylamine (40.5 mg, 0.400 mmol). The reaction mixture was stirred at60° C. for 3 hours. After gently blowing nitrogen over the reactionmixture to remove the solvent, the crude material was purified usingreverse phase HPLC using a mixture of water and acetonitrile as theeluent to give the desired product. ¹H NMR (400 MHz, DMSO-d₆) δ 9.49 (s,1H), 8.80 (s, 1H), 8.27 (d, J=9.0 Hz, 1H), 7.48 (dd, J=11.1, 8.3 Hz,1H), 7.37 (dd, J=8.2, 3.8 Hz, 1H), 6.27 (d, J=3.1 Hz, 1H), 6.06 (m, 1H),5.08 (m, 1H), 4.59 (q, J=6.6 Hz, 1H), 2.27 (s, 3H), 1.92 (m, 2H), 1.36(d, J=6.6 Hz, 3H), 0.92 (t, J=7.3 Hz, 3H). MS: (ES) m/z calculated forC₂₁H₂₀FN₃O₄ [M+Na]⁺ 420.1, found 420.4. The other diastereomer wasobtained similarly. ¹H NMR (400 MHz, DMSO-d₆) δ 9.48 (s, 1H), 8.80 (s,1H), 8.29 (d, J=8.1 Hz, 1H), 7.48 (dd, J=11.1, 8.2 Hz, 1H), 7.37 (dd,J=8.3, 3.8 Hz, 1H), 6.27 (d, J=3.1 Hz, 1H), 6.09-6.04 (m, 1H), 5.13-5.05(m, 1H), 4.65-4.54 (m, 1H), 2.27 (s, 3H), 1.92 (m, 2H), 1.36 (d, J=6.7Hz, 3H), 0.92 (t, J=7.3 Hz, 3H). MS: (ES) m/z calculated for C₂₁H₂₀FN₃O₄[M+Na]⁺ 420.1, found 420.3.

Example 5: Synthesis of(R)-3-((5-fluoro-1,1-dimethyl-3-oxoisoindolin-4-yl)amino)-4-((1-(5-methylfuran-2-yl)propyl)amino)cyclobut-3-ene-1,2-dione

Step a:

A mixture of methyl 2-bromo-5-fluorobenzoate (5.00 g, 21.5 mmol) andcopper cyanide (2.12 g, 23.6 mmol) in DMF was heated at 90° C. for 1day, then cooled down to room temperature, diluted with ethyl acetate(300 mL), and filtered. The filtrate was washed with brine (5×50 mL) andthen with sat. NaHCO₃ (50 mL). The organic layer was dried over MgSO₄,filtered, concentrated in vacuo. This product was used in the next stepwithout further purification. MS: (ES) m/z calculated for C₉H₆FNO₂[M+H]⁺ 180.0, found 180.0.

Step b:

To a stirred solution of methyl 2-cyano-5-fluorobenzoate (3.85 g, 21.5mmol) in tetrahydrofuran (30 mL) and water (3 mL) at 0° C. was addedlithium hydroxide monohydrate (1.11 g, 26.5 mmol). The reaction waswarmed up to rt and stirred for 1 h. Then the solvent was evaporated andthe residue was diluted with water (100 mL) and 2 M HCl (20 mL). Thesolid was collected by filtration and dried under vacuum to give thedesired product. MS: (ES) m/z calculated for C₈H₄FNO₂ [M+H]⁺ 166.0,found 166.0.

Step c:

To a stirred solution of 2-cyano-5-fluorobenzoic acid (1.70 g, 10.3mmol) in anhydrous tetrahydrofuran (105 mL) at −78° C. was added 1.6 Msolution of methyl lithium in ether (25.74 mL, 41.2 mmol) dropwise. Themixture was stirred at −78° C. for 1 h and was then slowly warmed up tort, quenched with saturated ammonium chloride, and extracted with ethylacetate. The organic layer was purified by silica gel chromatography(0-100% ethyl acetate/hexane) to give6-fluoro-3,3-dimethylisoindolin-1-one. MS: (ES) m/z calculated forC₁₀H₁₀FNO [M+H]⁺ 180.0, found 180.0.

Step d:

A reaction vial containing 6-fluoro-3,3-dimethylisoindolin-1-one (620mg, 3.46 mmol) in concentrated H₂SO₄ (1 mL) was cooled in an ice-bath. Amixture of concentrated H₂SO₄ (1 mL) with 70% HNO₃ (0.25 mL, 3.8 mmol)was added drop-wise and the reaction mixture was stirred at 0° C. for 2h then carefully quenched with ice and diluted to 10 mL with cold water.The solid was filtered, washed with water and dried under vacuum to givethe desired product 6-fluoro-3,3-dimethyl-7-nitroisoindolin-1-one. MS:(ES) m/z calculated for C₁₀H₉FN₂O₃ [M+H]⁺ 225.0, found 225.0.

Step e:

To a solution of 6-fluoro-3,3-dimethyl-7-nitroisoindolin-1-one (0.56 g,2.50 mmol) in ethanol (10 mL) and water (1 mL) at room temperature wasadded iron powder (0.58 g, 10.38 mmol) and ammonium chloride (1.90 g,34.6 mmol). The reaction mixture was warmed up to 90° C. and stirred for1 hour. Then it was cooled to room temperature, filtered through Celiteand rinsed with methanol (20 ml). The filtrate was concentrated todryness and the residue was diluted with ethyl acetate, washed withwater and brine. The combined organic layers were dried (Na₂SO₄),filtered, concentrated in vacuo, and purified by silica gelchromatography (0-100% ethyl acetate in hexanes) to provide the product7-amino-6-fluoro-3,3-dimethylisoindolin-1-one. MS: (ES) m/z calculatedfor C₁₀H₁₁FN₂O [M+H]⁺ 195.1, found 195.1.

Step f:

A mixture of 7-amino-6-fluoro-3,3-dimethylisoindolin-1-one (25 mg, 0.129mmol), 3,4-dimethoxycyclobut-3-ene-1,2-dione (22.0 mg, 0.155 mmol, 1.2equiv), and HCl solution in dioxane (4 M, 32.3 μL) in anhydrous MeOH(0.65 mL) was stirred at 60° C. for 18 hours. The solvents were removedto give the product which was directly used in the next step.

Step g:

A 20 mL vial was charged with3-((5-fluoro-1,1-dimethyl-3-oxoisoindolin-4-yl)amino)-4-methoxycyclobut-3-ene-1,2-dioneobtained above, followed by (1R)-1-(5-methyl-2-furyl)propan-1-aminetartrate salt (115.7 mg, 0.400 mmol) in methanol (0.5 mL) andtriethylamine (101 mg, 1.00 mmol). The reaction mixture was stirred at60° C. for 18 hours. After gently blowing nitrogen over the reactionmixture to remove the solvent, the crude material was purified usingreverse phase HPLC using a mixture of water and acetonitrile as theeluent.(R)-3-((5-fluoro-1,1-dimethyl-3-oxoisoindolin-4-yl)amino)-4-((1-(5-methylfuran-2-yl)propyl)amino)cyclobut-3-ene-1,2-dionewas obtained. ¹H NMR (400 MHz, DMSO-d₆) δ 9.48 (s, 1H), 8.81 (s, 1H),8.28 (d, J=9.1 Hz, 1H), 7.47 (dd, J=11.0, 8.2 Hz, 1H), 7.41 (dd, J=8.3,3.9 Hz, 1H), 6.27 (d, J=3.1 Hz, 1H), 6.09-6.03 (m, 1H), 5.08 (m, 1H),2.27 (s, 3H), 1.92 (m, 2H), 1.43 (s, 6H), 0.92 (t, J=7.3 Hz, 3H). MS:(ES) m/z calculated for C₂₂H₂₂FN₃O₄ [M+Na]⁺ 434.1, found 434.4.

Example 6: Synthesis of(R)-3-((5-fluoro-1,1,7-trimethyl-3-oxoisoindolin-4-yl)amino)-4-((1-(5-methylfuran-2-yl)propyl)amino)cyclobut-3-ene-1,2-dione

Step a:

To a solution of 7-amino-6-fluoro-3,3-dimethylisoindolin-1-one (150 mg,0.77 mmol) in AcOH (2 mL) in a water bath was added NIS (244 mg, 1.08mmol) in portions at room temperature. The resulting mixture was stirredin a water bath for 30 minutes, quenched with water (1 mL) and extractedwith ethyl acetate (10 mL). The organic layer was washed with brine (10mL) and then dried over MgSO₄. The solvent was evaporated under reducedpressure to give a brown solid, which was purified by silica gelchromatography (0-60% ethyl acetate in hexanes) to give7-amino-6-fluoro-4-iodo-3,3-dimethylisoindolin-1-one. MS: (ES) m/zcalculated for C₁₀H₁₀FIN₂O [M+H]⁺ 321.0, found 321.0.

Step b:

To a solution of 7-amino-6-fluoro-4-iodo-3,3-dimethylisoindolin-1-one(370 mg, 1.16 mmol) in dioxane (12 mL) was added CsF (705 mg, 4.64mmol), 2,4,6-trimethyl-1,3,5,2,4,6-trioxatriborinane (435 mg, 3.47mmol), and [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II)(95 mg, 0.116 mmol). The resulting mixture was stirred at 80° C.overnight. The reaction was partitioned between water (20 mL) and ethylacetate (30 mL). The organic layer was washed with brine (20 mL) andthen dried over MgSO₄, filtered, and concentrated to give the crude,which was purified by silica gel chromatography (0-80% ethyl acetate inhexanes) to give 7-amino-6-fluoro-3,3,4-trimethylisoindolin-1-one. MS:(ES) m/z calculated for C₁₁H₁₃FN₂O [M+H]⁺ 209.1, found 209.1.

Step c:

A mixture of 7-amino-6-fluoro-3,3,4-trimethylisoindolin-1-one (40 mg,0.192 mmol), 3,4-dimethoxycyclobut-3-ene-1,2-dione (32.8 mg, 0.231 mmol,1.2 equiv), and HCl solution in dioxane (4 M, 48 μL) in anhydrous MeOH(1.0 mL) was stirred at 60° C. for 18 hours. The solvents were removedto give the product which was directly used in the next step.

Step d:

A 20 mL vial was charged with3-((5-fluoro-1,1,7-trimethyl-3-oxoisoindolin-4-yl)amino)-4-methoxycyclobut-3-ene-1,2-dioneobtained above, followed by (JR)-1-(5-methyl-2-furyl)propan-1-aminetartrate salt (222.7 mg, 0.77 mmol) in methanol (1.0 mL) andtriethylamine (233 mg, 2.3 mmol). The reaction mixture was stirred at60° C. for 18 hours. After gently blowing nitrogen over the reactionmixture to remove the solvent, the crude material was purified usingreverse phase HPLC using a mixture of water and acetonitrile as theeluent.(R)-3-((5-fluoro-1,1,7-trimethyl-3-oxoisoindolin-4-yl)amino)-4-((1-(5-methylfuran-2-yl)propyl)amino)cyclobut-3-ene-1,2-dionewas obtained. ¹H NMR (400 MHz, DMSO-d₆) δ 9.41 (s, 1H), 8.83 (s, 1H),8.31 (d, J=9.1 Hz, 1H), 7.29 (d, J=12.0 Hz, 1H), 6.26 (d, J=3.1 Hz, 1H),6.12-6.02 (m, 1H), 5.08 (m, 1H), 2.41 (s, 3H), 2.27 (s, 3H), 1.90 (m,2H), 1.49 (s, 6H), 0.92 (t, J=7.3 Hz, 3H). MS: (ES) m/z calculated forC₂₃H₂₄FN₃O₄ [M+Na]⁺448.2, found 448.4.

Example 7: Synthesis of(R)-2-(4-chloro-7-((2-((1-(5-methylfuran-2-yl)propyl)amino)-3,4-dioxocyclobut-1-en-1-yl)amino)-1-oxoisoindolin-2-yl)-4-methoxybenzoicAcid

Step a:

A 1 L round bottom flask containing 4-chloroisoindolin-1-one (25.0 g,0.149 mole) in concentrated H₂SO₄ (50 mL) was cooled in an ice-bath. Amixture of concentrated H₂SO₄ (50 mL) with 70% HNO₃ (10 mL, 0.16 mole,1.05 equiv.) was added drop-wise and the reaction mixture was stirred at0° C. for 2 h then carefully quenched with ice and diluted to 1 L withcold water. The solid was filtered, washed with water and dried underhigh vacuum to afford 4-chloro-7-nitro-isoindolin-1-one. MS: (ES) m/zcalculated for C₈H₅ClN₂O₃ [M−H]⁻ 212.0, found 212.0.

Step b:

To a stirred mixture of 4-chloro-7-nitro-isoindolin-1-one (23 g, 108mmol) in ethanol at room temperature was added iron powder (18.2 g, 324mmol), followed by 4 M HCl in dioxane (162 mL, 648 mmol). The reactionmixture was stirred at room temperature for 1 h then concentrated invacuo. The residue was diluted with ethyl acetate and neutralized withsaturated sodium bicarbonate solution and extracted with ethyl acetate(2×500 mL). The combined organic layers were dried (Na₂SO₄), filtered,and concentrated in vacuo to afford 7-amino-4-chloro-isoindolin-1-one.MS: (ES) m/z calculated for C₈H₇ClN₂O [M+H]⁺ 183.2, found 183.2.

Step c:

To a reaction vial containing 7-amino-4-chloro-isoindolin-1-one (250 mg,1.37 mmol) in dioxane (10 mL) was added methyl2-bromo-5-methoxy-benzoate (502 mg, 2.05 mmol), cesium carbonate (893mg, 2.74 mmol), copper iodide (104 mg, 0.55 mmol) and(1S,2S)—N1,N2-dimethylcyclohexane-1,2-diamine (156 mg, 1.1 mmol). Themixture was purged with nitrogen, then warmed to 110° C. The reactionwas stirred at 110° C. for 1 h and the reaction was monitored by LC-MS.Following completion, the reaction was allowed to cool and was thenfiltered through Celite and rinsed with ethyl acetate. The crude waspurified by silica gel chromatography (0-50% ethyl acetate/hexane) togive methyl2-(7-amino-4-chloro-1-oxo-isoindolin-2-yl)-5-methoxy-benzoate. MS: (ES)m/z calculated for C₁₇H₁₅ClN₂O₄[M+H]⁺ 347.1, found 347.1.

Step d:

A mixture of methyl2-(7-amino-4-chloro-1-oxo-isoindolin-2-yl)-5-methoxy-benzoate (160 mg,0.46 mmol) and 3,4-dimethoxycyclobutane-1,2-dione (131 mg, 0.92 mmol) inanhydrous methanol (5 mL) was stirred at 60° C. overnight. The reactionmixture was evaporated and the residue was stirred in ethyl acetate (5mL) at 50° C. for 30 min, then allowed to cool to room temperature. Themixture was filtered and dried to give the product methyl2-[4-chloro-7-[(2-methoxy-3,4-dioxo-cyclobutyl)amino]-1-oxo-isoindolin-2-yl]-5-methoxy-benzoate.MS: (ES) m/z calculated for C₂₂H₁₇ClN₂O₇ [M+H]⁺ 457.1, found 457.1.

Step e:

To methyl2-(4-chloro-7-((2-methoxy-3,4-dioxocyclobut-1-en-1-yl)amino)-1-oxoisoindolin-2-yl)-4-methoxybenzoate(60 mg, 0.12 mmol) was added (R)-1-(5-methylfuran-2-yl)propan-1-amine(22 mg, 0.15 mmol) in methanol (5 mL) at ambient temperature. Thereaction mixture was stirred overnight at 60° C. and then concentrated.The residue was diluted with dichloromethane and washed with water. Theorganic layer was concentrated. To the residue was added THF (5 mL),water (1 mL), and LiOH (large excess). The reaction mixture was stirredat room temperature for 1 h and 60° C. for 2 h. Aqueous HCl (1N) anddichloromethane were added and the combined organic layer wasconcentrated. THF was added and the mixture was filtered to get rid ofthe solid. The residue was purified by reverse phase HPLC using amixture of water and acetonitrile as the eluent.(R)-2-(4-chloro-7-((2-((1-(5-methylfuran-2-yl)propyl)amino)-3,4-dioxocyclobut-1-en-1-yl)amino)-1-oxoisoindolin-2-yl)-4-methoxybenzoicacid was obtained. ¹H NMR (400 MHz, Methanol-d₄) δ 8.12-8.03 (m, 2H),7.59 (d, J=8.8 Hz, 1H), 7.15-7.02 (m, 2H), 6.20 (s, 1H), 5.94 (s, 1H),5.20-4.80 (m, 3H), 3.90 (s, 3H), 2.24 (s, 3H), 2.03-1.85 (m, 2H), 0.99(t, J=7.4 Hz, 3H). MS: (ES) m/z calculated for C₂₈H₂₄ClN₃O₇ [M+H]⁺550.1, found 550.3.

Example 8: Synthesis of(R)-3-((5-fluoro-3-oxoisoindolin-4-yl)amino)-4-((1-(5-methylfuran-2-yl)propyl-1-d)amino)cyclobut-3-ene-1,2-dione

Step a:

To a 40 mL vial was added (R)-2-methylpropane-2-sulfinamide (1.83 g,15.1 mmol), 1-(5-methylfuran-2-yl)propan-1-one (2.0 mL, 15.1 mmol), andtitanium ethoxide (7.8 mL). The reaction mixture was stirred at 60° C.overnight. The reaction was diluted with dichloromethane (100 mL) andquenched with sodium sulfate decahydrate (10.2 g). The reaction mixturewas filtered through celite and rinsed with dichloromethane. Evaporationof the solvent gave the crude product which was purified by silica gelchromatography using a mixture of ethyl acetate and hexane as theeluent.

Step b:

To a mixture of the product from the previous step (1122 mg, 4.65 mmol),THF (17.5 mL), and D₂O (0.36 mL) at −55° C. was added sodiumborodeuteride (575 mg, 13.7 mmol). The reaction mixture was stirred atthe same temperature for 3 h. After gently blowing nitrogen over thereaction mixture to remove the solvent, dichloromethane was added. Themixture was filtered and the filtrate was concentrated. The residue waspurified by silica gel chromatography using a mixture of dichloromethaneand methyl tert-butyl ether as the eluent.

Step c:

The product from the previous step (422 mg, 1.73 mmol) was dissolved inMeOH (5 mL) at 0° C., and acetyl chloride (400 μL) was added. Thereaction was stirred for 1 h. After gently blowing nitrogen over thereaction mixture to remove the solvent, a mixture of methyl tert-butylether and water was added. The aqueous layer was collected and basifiedwith sodium carbonate solution (2 M). The mixture was extracted withCHCl₃ (3×). The combined organic layers were dried over Na₂SO₄,filtered, and concentrated to give the product.

Step d:

To3-((5-fluoro-3-oxoisoindolin-4-yl)amino)-4-methoxycyclobut-3-ene-1,2-dione(41.4 mg, 0.150 mmol) was added(R)-1-(5-methylfuran-2-yl)propan-1-d-1-amine (31.9 mg, 0.228 mmol) inmethanol (1 mL) at ambient temperature. The reaction mixture was stirredovernight at 45° C. After gently blowing nitrogen over the reactionmixture to remove the solvent, a mixture of aqueous HCl (1N) anddichloromethane was added. The mixture was filtered and the residue waspurified by prep TLC using ethyl acetate as the eluent.(R)-3-((5-fluoro-3-oxoisoindolin-4-yl)amino)-4-((1-(5-methylfuran-2-yl)propyl-1-d)amino)cyclobut-3-ene-1,2-dionewas obtained. ¹H NMR (400 MHz, DMSO-d₆) (9.47 (s, 1H), 8.70 (s, 1H),8.25 (s, 1H), 7.46 (dd, J=11.2, 8.2 Hz, 1H), 7.33 (dd, J=8.2, 3.7 Hz,1H), 6.25 (d, J=3.1 Hz, 1H), 6.06-6.03 (m, 1H), 4.32 (s, 2H), 2.26 (s,3H), 2.00-1.80 (m, 2H), 0.90 (t, J=7.3 Hz, 3H). MS: (ES) m/z calculatedfor C₂₀DH₁₇FN₃O₄ [M+H]⁺ 385.1, found 385.4.

Example 9: Synthesis of(R)-3-((5-fluoro-3-oxoisoindolin-4-yl)amino)-4-((1-(5-methyloxazol-2-yl)propyl)amino)cyclobut-3-ene-1,2-dione

Step a:

To a 40 mL vial was added (R)-2-((tert-butoxycarbonyl)amino)butanoicacid (1.00 g, 4.92 mmol),1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate (HATU, 1.96 g, 5.16 mmol), DMF (6 mL), andtriethylamine (1.51 mL). The reaction mixture was stirred for 2 min andprop-2-yn-1-amine (378 μL, 5.90 mmol) was added. The reaction mixturewas further stirred at room temperature for 2 h. The reaction mixturewas diluted with water and diethyl ether, and extracted with diethylether (6×). The combined organic layers were dried over Na₂SO₄,filtered, and concentrated. The residue was purified by silica gelchromatography using a mixture of ethyl acetate and dichloromethane asthe eluent.

Step b:

To a 40 mL vial were added auric chloride (128 mg, 0.424 mmol),acetonitrile (10 mL), and the product from the previous step (501 mg,2.09 mmol). The reaction mixture was stirred at 50° C. overnight. Aftergently blowing nitrogen over the reaction mixture to remove the solvent,a mixture of saturated ammonium chloride solution and methyl tert-butylether was added. The mixture was extracted with methyl tert-butyl ether(3×). The combined organic layers were dried over Na₂SO₄, filtered, andconcentrated. The residue was purified by silica gel chromatographyusing a mixture of ethyl acetate and hexane as the eluent.

Step c:

To a stirred solution of the product from the previous step (301 mg,1.25 mmol) in dichloromethane (1 mL) was added 4N HCl in dioxane (1 mL).The mixture was stirred at room temperature overnight. After gentlyblowing nitrogen over the reaction mixture to remove the solvent, amixture of water and hexane was added. The aqueous layer was collectedand basified with saturated sodium bicarbonate solution. The mixture wasextracted with dichloromethane (3×) and chloroform/isopropanol (2/1,v/v) (2×). The combined organic layers were dried over Na₂SO₄, filtered,and concentrated to give the product.

Step d:

To3-((5-fluoro-3-oxoisoindolin-4-yl)amino)-4-methoxycyclobut-3-ene-1,2-dione(42.4 mg, 0.154 mmol) was added(R)-1-(5-methyloxazol-2-yl)propan-1-amine (25.9 mg, 0.185 mmol) inmethanol (0.5 mL) at ambient temperature. The reaction mixture wasstirred overnight at 50° C. After gently blowing nitrogen over thereaction mixture to remove the solvent, a mixture of aqueous HCl (1N)and dichloromethane was added. The mixture was filtered and the residuewas purified by prep TLC using a mixture of dichloromethane andacetonitrile (1/1, v/v) as the eluent.(R)-3-((5-fluoro-3-oxoisoindolin-4-yl)amino)-4-((1-(5-methyloxazol-2-yl)propyl)amino)cyclobut-3-ene-1,2-dionewas obtained. ¹H NMR (400 MHz, DMSO-d₆) δ 9.63 (s, 1H), 8.73 (s, 1H),8.48 (d, J=7.5 Hz, 1H), 7.49 (dd, J=11.2, 8.2 Hz, 1H), 7.36 (dd, J=8.3,3.8 Hz, 1H), 6.87 (d, J=1.4 Hz, 1H), 5.33-5.20 (m, 1H), 4.34 (s, 2H),2.32 (d, J=1.2 Hz, 3H), 2.09-1.87 (m, 2H), 0.93 (t, J=7.4 Hz, 3H). MS:(ES) m/z calculated for C₁₉H₁₇FN₄O₄ [M+H]⁺ 385.1, found 385.4.

Example 10: Synthesis of(R)-2-(4-chloro-7-((2-((1-(5-methylfuran-2-yl)propyl)amino)-3,4-dioxocyclobut-1-en-1-yl)amino)-1-oxoisoindolin-2-yl)acetamide

Step a:

A 4 L Erlenmeyer flask containing 3-chloro-2-methyl-benzoic acid (100.0g, 0.586 mole) in concentrated H₂SO₄ (500 mL) was cooled in an ice-bath.70% HNO₃ (45.2 mL, 0.703 mole, 1.2 equiv.) was added drop-wise andreaction mixture was stirred at 0° C. for 2 h, then carefully quenchedwith ice and diluted to 4 L with cold water. A white solid was filtered,washed with water and dried under high vacuum to afford a mixture of3-chloro-2-methyl-6-nitro-benzoic acid and3-chloro-2-methyl-5-nitro-benzoic acid in 3:1 ratio. MS: (ES) m/zcalculated for C₈H₅ClNO₄ [M−H]⁻ 214.0, found 214.0.

Step b:

The mixture of isomeric acids from the previous step (50 g, 232.0 mmol)was dissolved in anhydrous DMF (200 mL), anhydrous Na₂CO₃ (27.0 g, 255.2mmol, 1.1 equiv.) was added, and the reaction was stirred at roomtemperature for 30 minutes. Methyl iodide (15.9 mL, 255.2 mmol, 1.1equiv.) was added and stirring was continued at room temperature for 3h. The reaction mixture was diluted with water (1.2 L) and the productwas extracted using Et₂O (3×250 mL). The combined organic layers werewashed with brine (4×100 mL), dried over MgSO₄, filtered and evaporatedto give product.

Step c:

The mixture of isomeric esters from the previous step (49.7 g, 216.5mmol) was dissolved in CCl₄ (400 mL) and N-bromosuccinimide (57.8 g,324.7 mmol, 1.5 equiv.) was added followed by benzoyl peroxide (10.4 g,43.2 mmol, 0.20 equiv). The reaction mixture was stirred under refluxovernight then cooled to room temperature and filtered. The filtrate wasevaporated and the residue was purified by silica gel chromatography(100:0 to 9:1 Hex:EtOAc) to give product as a single isomer. ¹H NMR (400MHz, CDCl₃) δ 8.07 (d, J=9.2 Hz, 1H), 7.65 (d, J=9.2 Hz, 1H), 4.63 (s,2H), 4.01 (s, 3H).

Step d:

A mixture of the product from the previous step (316 mg, 1.02 mmol),2-aminoacetamide hydrochloride (171 mg, 1.55 mmol) and triethylamine(427 μL, 3.06 mmol) in ethanol (2 mL) was stirred at room temperatureovernight. After gently blowing nitrogen over the reaction mixture toremove the solvent, a mixture of aqueous HCl (1N) and water was added.The mixture was filtered and the residue was rinsed with aqueous HCl(1N) and water to afford the desired product.

Step e:

To a stirred mixture of the product from the previous step (249 mg,0.925 mmol) in ethanol (1 mL) at room temperature was added iron powder(158 mg, 2.68 mmol), followed by 4 M HCl in dioxane (0.46 mL, 1.84mmol). The reaction mixture was stirred at room temperature overnight.Saturated sodium bicarbonate aqueous solution was added and the mixturewas diluted with methanol. The mixture was filtered through celite andrinsed with methanol. The filtrate was concentrated to remove methanoland subsequently extracted with dichloromethane. The combined organiclayers were concentrated to afford the desired product.

Step f:

A mixture of the product from the previous step (131 mg, 0.55 mmol) and3,4-dimethoxycyclobutane-1,2-dione (118 mg, 0.83 mmol) in anhydrousmethanol (2 mL) was stirred at 60° C. overnight. After gently blowingnitrogen over the reaction mixture to remove the solvent, ethyl acetatewas added. The mixture was filtered, rinsed with ethyl acetate, anddried to give the product.

Step g:

To2-(4-chloro-7-((2-methoxy-3,4-dioxocyclobut-1-en-1-yl)amino)-1-oxoisoindolin-2-yl)acetamide(39.9 mg, 0.114 mmol) was added (R)-1-(5-methylfuran-2-yl)propan-1-amine(23.0 mg, 0.165 mmol) in ethanol (1 mL) at ambient temperature. Thereaction mixture was stirred overnight at 65° C. After gently blowingnitrogen over the reaction mixture to remove the solvent, a mixture ofaqueous HCl (1N) and dichloromethane was added. The mixture was filteredand the residue was rinsed with water, dichloromethane, and methyltert-butyl ether.(R)-2-(4-chloro-7-((2-((1-(5-methylfuran-2-yl)propyl)amino)-3,4-dioxocyclobut-1-en-1-yl)amino)-1-oxoisoindolin-2-yl)acetamidewas obtained. ¹H NMR (400 MHz, DMSO-d₆) δ 9.74 (s, 1H), 9.12 (d, J=8.9Hz, 1H), 7.67-7.52 (m, 3H), 7.27-7.18 (m, 1H), 6.25 (d, J=3.1 Hz, 1H),6.06-6.02 (m, 1H), 5.18-5.08 (m, 1H), 4.48 (s, 2H), 4.11 (s, 2H), 2.25(s, 3H), 2.01-1.80 (m, 2H), 0.90 (t, J=7.3 Hz, 3H). MS: (ES) m/zcalculated for C₂₂H₂₁ClN₄O₅ [M+Na]⁺479.1, found 479.0.

The following compounds were made using similar synthetic methods asdescribed herein with the appropriate reagents and were characterized byMS (Mass spectrometry) and NMR as illustrated in Table 1.

TABLE 1 Characterization of compounds Structure NMR MS

1H NMR (400 MHz, Methanol- d4) δ 8.06 (d, J = 8.6 Hz, 1H), 7.67 (d, J =8.7 Hz, 1H), 7.56 (d, J = 8.7 Hz, 1H), 7.11 (d, J = 2.5 Hz, 1H), 7.02(dd, J = 8.6, 2.5 Hz, 1H), 6.19 (s, 1H), 5.95-5.91 (m, 1H), 5.28-5.20(m, 1H), 4.93- 4.90 (m, 2H), 3.88 (s, 3H), 2.23 (s, 3H), 2.08-1.88 (m,2H), 0.99 (t, J = 7.4 Hz, 3H). MS: (ES) m/z calculated for C28H25ClN4O6[M + H]+ 549.2, found 549.3.

1H NMR (400 MHz, DMSO-d6) δ 11.89 (s, 1H), 9.82 (s, 1H), 9.14 (d, J =8.9 Hz, 1H), 7.69-7.53 (m, 3H), 7.32-7.16 (m, 2H), 6.26 (d, J = 3.1 Hz,1H), 6.06- 6.03 (m, 1H), 5.20-5.12 (m, 1H), 4.97 (s, 2H), 2.26 (s, 3H),2.06 (s, 3H), 2.02-1.83 (m, 2H), 0.93 (t, J = 7.3 Hz, 3H). MS: (ES) m/zcalculated for C26H24N4O5 [M + Na]+ 495.2, found 495.4.

1H NMR (400 MHz, Methanol- d4) δ 7.94 (d, J = 7.8 Hz, 1H), 7.50-7.31 (m,4H), 6.19 (d, J = 3.2 Hz, 1H), 5.96-5.93 (m, 1H), 5.22-5.12 (m, 1H),4.83 (s, 2H), 2.45 (s, 3H), 2.25 (s, 3H), 2.06- 1.86 (m, 2H), 0.99 (t, J= 7.4 Hz, 3H). MS: (ES) m/z calculated for C28H24FN3O6 [M − H]− 516.2,found 516.3.

1H NMR (400 MHz, DMSO-d6) δ 9.81 (s, 1H), 9.14 (d, J = 8.9 Hz, 1H), 7.88(d, J = 9.1 Hz, 3H), 7.72- 7.55 (m, 2H), 7.25 (d, J = 7.1 Hz, 1H), 6.45(d, J = 10.1 Hz, 1H), 6.27 (d, J = 3.1 Hz, 1H), 6.07- 6.04 (m, 1H),5.20-5.12 (m, 1H), 4.89 (s, 2H), 2.27 (d, J = 1.1 Hz, 3H), 2.06-1.80 (m,2H), 0.93 (t, J = 7.3 Hz, 3H). MS: (ES) m/z calculated for C25H22N4O5[M + H]+ 459.2, found 459.4.

1H NMR (400 MHz, Methanol- d4) δ 8.10-7.99 (m, 1H), 7.55- 7.37 (m, 2H),7.14-6.99 (m, 2H), 6.19 (d, J = 3.2 Hz, 1H), 5.96- 5.93 (m, 1H),5.21-5.11 (m, 1H), 3.89 (s, 3H), 4.83 (s, 2H), 2.25 (s, 3H), 2.07-1.88(m, 2H), 0.99 (t, J = 7.4 Hz, 3H). MS: (ES) m/z calculated forC28H24FN3O7 [M + Na]+ 556.2, found 556.5.

1H NMR (400 MHz, DMSO-d6) δ 9.27 (s, 1H), 8.75 (s, 1H), 8.11 (d, J = 7.7Hz, 1H), 7.25 (d, J = 12.6 Hz, 1H), 6.26 (d, J = 3.1 Hz, 1H), 6.07-6.04(m, 1H), 5.12-5.03 (m, 1H), 4.24 (s, 2H), 3.88 (s, 3H), 2.27 (s, 3H),1.99-1.82 (m, 2H), 0.92 (t, J = 7.3 Hz, 3H). MS: (ES) m/z calculated forC21H20FN3O5 [M + Na]+ 436.2, found 436.4.

1H NMR (400 MHz, DMSO-d6) δ 9.17 (s, 1H), 8.64 (s, 1H), 7.98 (d, J = 9.1Hz, 1H), 7.25 (d, J = 10.5 Hz, 1H), 6.25 (d, J = 3.1 Hz, 1H), 6.07-6.04(m, 1H), 5.11-5.03 (m, 1H), 4.33 (s, 2H), 3.82 (s, 3H), 2.28 (s, 3H),1.99-1.82 (m, 2H), 0.92 (t, J = 7.3 Hz, 3H). MS: (ES) m/z calculated forC21H20FN3O5 [M + Na]+ 436.2, found 436.0.

1H NMR (400 MHz, Methanol- d4) δ 8.14-8.08 (m, 1H), 7.27- 7.23 (, 1H),6.25 (d, J = 3.1 Hz, 1H), 6.00-5.96 (m, 1H), 5.20- 5.05 (m, 1H), 4.61(s, 2H), 4.33 (s, 2H), 2.26 (s, 3H), 2.15-1.90 (m, 2H), 1.02 (t, J = 7.4Hz, 3H). MS: (ES) m/z calculated for C22H21FN4O5 [M + H]+ 441.2, found441.4.

1H NMR (400 MHz, DMSO-d6) δ 9.76 (s, 1H), 9.15 (d, J = 8.8 Hz, 1H),7.70-7.63 (m, 2H), 7.52 (d, J = 8.7 Hz, 1H), 7.39 (d, J = 3.0 Hz, 1H),7.25 (dd, J = 8.7, 3.1 Hz, 1H), 6.23 (d, J = 3.2 Hz, 1H), 6.04- 6.00 (m,1H), 5.17-5.08 (m, 1H), 4.79 (s, 2H), 3.83 (s, 3H), 2.23 (s, 3H),2.06-1.78 (m, 2H), 0.89 (t, J = 7.3 Hz, 3H). MS: (ES) m/z calculated forC28H24ClN3O7 [M + H]+ 550.1, found 550.3.

1H NMR (400 MHz, Methanol- d4) δ 8.12-8.05 (m, 1H), 7.95 (d, J = 7.8 Hz,1H), 7.58 (d, J = 8.8 Hz, 1H), 7.39-7.33 (m, 2H), 6.22-6.18 (m, 1H),5.95-5.91 (m, 1H), 5.20-4.80 (m, 3H), 2.46 (s, 3H), 2.24 (s, 3H), 2.12-1.82 (m, 2H), 0.99 (t, J = 7.4 Hz, 3H). MS: (ES) m/z calculated forC28H24ClN3O6 [M + H]+ 534.1, found 534.3.

1H NMR (400 MHz, DMSO-d6) δ 9.78 (s, 1H), 9.14 (d, J = 9.1 Hz, 1H),7.78-7.20 (m, 9H), 6.23 (d, J = 3.1 Hz, 1H), 6.05-6.00 (m, 1H),5.20-5.10 (m, 1H), 4.95- 4.83 (m, 2H), 2.23 (s, 3H), 2.03- 1.77 (m, 2H),0.90 (t, J = 7.3 Hz, 3H). MS: (ES) m/z calculated for C27H24N4O5 [M +Na]+ 507.2, found 507.5.

1H NMR (400 MHz, Methanol- d4) δ 7.48 (d, J = 7.7 Hz, 1H), 7.34 (d, J =7.8 Hz, 1H), 6.21 (d, J = 3.1 Hz, 1H), 5.98-5.95 (m, 1H), 5.20-5.10 (m,1H), 4.37 (s, 2H), 2.37 (s, 3H), 2.28 (d, J = 1.0 Hz, 3H), 2.08-1.88 (m,2H), 1.01 (t, J = 7.4 Hz, 3H). MS: (ES) m/z calculated for C21H21N3O4 [M− H]− 378.1, found 378.3.

1H NMR (400 MHz, DMSO-d6) δ 12.08 (s, 1H), 9.80 (s, 1H), 9.11 (d, J =9.0 Hz, 1H), 7.71 (dd, J = 7.1, 2.0 Hz, 1H), 7.64-7.53 (m, 2H),7.44-7.38 (m, 1H), 7.24 (d, J = 7.0 Hz, 1H), 6.30 (d, J = 6.8 Hz, 1H),6.24 (d, J = 3.1 Hz, 1H), 6.04-6.01 (m, 1H), 5.20-5.10 (m, 1H), 4.93 (s,2H), 2.24 (s, 3H), 2.00-1.77 (m, 2H), 0.91 (t, J = 7.4 Hz, 3H). MS: (ES)m/z calculated for C25H22N4O5 [M + Na]+ 481.2, found 481.4.

1H NMR (400 MHz, Chloroform- d) δ 7.10-6.90 (m, 2H), 6.15 (d, J = 3.1Hz, 1H), 5.89-5.85 (m, 1H), 5.25-5.15 (m, 1H), 4.19 (s, 2H), 3.71 (s,3H), 2.22 (s, 3H), 1.85-1.60 (m, 2H), 0.97-0.85 (m, 3H). MS: (ES) m/zcalculated for C21H21N3O5 [M − H]− 394.1, found 394.3.

1H NMR (400 MHz, DMSO-d6) δ 9.38 (s, 1H), 8.70 (s, 1H), 8.18 (d, J = 8.9Hz, 1H), 7.31 (d, J = 11.8 Hz, 1H), 6.25 (d, J = 3.1 Hz, 1H), 6.06-6.02(m, 1H), 5.12-5.00 (m, 1H), 4.26 (s, 2H), 2.25 (s, 6H), 1.98-1.80 (m,2H), 0.90 (t, J = 7.3 Hz, 3H). MS: (ES) m/z calculated for C21H20FN3O4[M + H]+ 398.1, found 398.4.

1H NMR (400 MHz, DMSO-d6) δ 9.70 (s, 1H), 8.73 (s, 1H), 8.52 (d, J = 9.7Hz, 1H), 7.49 (dd, J = 11.2, 8.2 Hz, 1H), 7.36 (dd, J = 8.3, 3.9 Hz,1H), 6.88 (d, J = 1.5 Hz, 1H), 5.23-5.15 (m, 1H), 4.34 (s, 2H), 2.32 (d,J = 1.2 Hz, 3H), 2.29- 2.21 (m, 1H), 0.95 (d, J = 6.8 Hz, 3H), 0.92 (d,J = 6.7 Hz, 3H). MS: (ES) m/z calculated for C20H19FN4O4 [M + H]+ 399.1,found 399.4.

1H NMR (400 MHz, DMSO-d6) δ 10.62 (s, 1H), 9.04 (s, 1H), 8.55 (s, 1H),7.94-7.84 (m, 1H), 6.88 (d, J = 9.9 Hz, 1H), 6.25 (d, J = 3.1 Hz, 1H),6.07-6.04 (m, 1H), 5.13-5.01 (m, 1H), 4.28 (s, 2H), 2.27 (d, J = 1.0 Hz,3H), 1.99- 1.81 (m, 2H), 0.92 (t, J = 7.3 Hz, 3H). MS: (ES) m/zcalculated for C20H18FN3O5 [M + H]+ 400.1, found 400.4.

1H NMR (400 MHz, DMSO-d6) δ 9.49 (s, 1H), 8.91 (s, 1H), 8.24 (d, J = 9.7Hz, 1H), 7.61 (dd, J = 10.9, 8.6 Hz, 1H), 6.21 (d, J = 3.1 Hz, 1H),6.05-6.02 (m, 1H), 5.00- 4.91 (m, 1H), 4.41 (s, 2H), 2.26 (s, 3H),2.21-2.11 (m, 1H), 0.95 (d, J = 6.7 Hz, 3H), 0.87 (d, J = 6.7 Hz, 3H).MS: (ES) m/z calculated for C21H19F2N3O4 [M + H]+ 416.1, found 416.4.

1H NMR (400 MHz, DMSO-d6) δ 9.59 (s, 1H), 8.93 (s, 1H), 8.30 (d, J = 9.8Hz, 1H), 7.77 (d, J = 10.6 Hz, 1H), 6.21 (d, J = 3.1 Hz, 1H), 6.04 (d, J= 3.0 Hz, 1H), 5.01- 4.93 (m, 1H), 4.32 (s, 2H), 2.26 (d, J = 1.2 Hz,3H), 2.23-2.13 (m, 1H), 0.95 (d, J = 6.7 Hz, 3H), 0.87 (d, J = 6.7 Hz,3H). MS: (ES) m/z calculated for C21H19ClFN3O4 [M + H]+ 432.1, found432.4.

1H NMR (400 MHz, DMSO-d6) δ 9.50 (s, 1H), 8.82 (s, 1H), 8.08- 8.00 (m,2H), 7.93 (s, 1H), 6.27 (d, J = 3.1 Hz, 1H), 6.08-6.05 (m, 1H),5.13-5.02 (m, 1H), 4.33 (s, 2H), 2.28 (s, 3H), 2.00- 1.80 (m, 2H), 0.92(t, J = 7.3 Hz, 3H). MS: (ES) m/z calculated for C20H17Cl2N3O4 [M + H]+434.1, found 434.3.

1H NMR (400 MHz, DMSO-d6) δ 9.83 (s, 1H), 9.15 (d, J = 9.5 Hz, 1H),7.66-7.55 (m, 3H), 7.29- 7.20 (m, 1H), 6.23 (d, J = 3.1 Hz, 1H),6.07-6.03 (m, 1H), 5.08- 5.00 (m, 1H), 4.50 (s, 2H), 4.14 (s, 2H), 2.27(s, 3H), 2.25-2.13 (m, 1H), 0.97 (d, J = 6.7 Hz, 3H), 0.88 (d, J = 6.7Hz, 3H). MS: (ES) m/z calculated for C23H23ClN4O5 [M + Na]+ 493.1, found493.4.

1H NMR (400 MHz, DMSO-d6) δ 9.49 (s, 1H), 8.56 (s, 1H), 8.08 (d, J = 9.8Hz, 1H), 7.68 (d, J = 8.0 Hz, 1H), 7.38 (d, J = 8.1 Hz, 1H), 6.22 (d, J= 3.1 Hz, 1H), 6.06- 6.02 (m, 1H), 5.00-4.92 (m, 1H), 4.32 (s, 2H), 2.27(s, 3H), 2.24-2.09 (m, 1H), 0.96 (d, J = 6.7 Hz, 3H), 0.87 (d, J = 6.7Hz, 3H). MS: (ES) m/z calculated for C21H20ClN3O4 [M + H]+ 414.1, found414.4.

1H NMR (400 MHz, DMSO-d6) δ 11.55 (s, 1H), 9.82 (s, 1H), 8.16- 8.00 (m,2H), 6.30-6.22 (m, 1H), 6.07-6.02 (m, 1H), 5.14- 5.02 (m, 1H), 2.25 (s,3H), 2.04- 1.75 (m, 2H), 0.91 (t, J = 6.5 Hz, 3H). MS: (ES) m/zcalculated for C20H15Cl2N3O5 [M + Na]+ 470.0, found 470.2.

1H NMR (400 MHz, DMSO-d6) δ 9.53 (s, 1H), 8.93 (s, 1H), 8.23 (d, J = 8.7Hz, 1H), 7.76 (d, J = 10.6 Hz, 1H), 6.25 (d, J = 3.1 Hz, 1H), 6.06-6.02(m, 1H), 5.12-5.00 (m, 1H), 4.32 (s, 2H), 2.25 (s, 3H), 1.99-1.80 (m,2H), 0.90 (t, J = 7.3 Hz, 3H). MS: (ES) m/z calculated for C20H17ClFN3O4[M + H]+ 418.1, found 418.4.

1H NMR (400 MHz, DMSO-d6) δ 9.43 (s, 1H), 8.57 (s, 1H), 8.08- 7.98 (m,1H), 7.68 (d, J = 8.0 Hz, 1H), 7.38 (d, J = 8.1 Hz, 1H), 6.26 (d, J =3.1 Hz, 1H), 6.06-6.03 (m, 1H), 5.12-5.00 (m, 1H), 4.31 (s, 2H), 2.26(s, 3H), 1.99- 1.80 (m, 2H), 0.91 (t, J = 7.3 Hz, 3H). MS: (ES) m/zcalculated for C20H18ClN3O4 [M + Na]+ 422.1, found 422.0.

1H NMR (400 MHz, DMSO-d6) δ 9.85 (s, 1H), 9.35 (d, J = 8.9 Hz, 1H), 8.93(s, 1H), 7.63-7.55 (m, 2H), 6.84 (s, 1H), 5.35-5.25 (m, 1H), 4.36 (s,2H), 2.29 (s, 3H), 2.07-1.85 (m, 2H), 0.92 (t, J = 7.3 Hz, 3H). MS: (ES)m/z calculated for C19H17ClN4O4 [M + H]+ 401.1, found 401.0.

1H NMR (400 MHz, DMSO-d6) δ 10.01 (s, 1H), 9.80 (d, J = 9.4 Hz, 1H),8.96 (s, 1H), 7.61 (d, J = 8.7 Hz, 1H), 7.52 (d, J = 8.7 Hz, 1H), 6.65(d, J = 3.2 Hz, 1H), 6.19 (d, J = 3.2 Hz, 1H), 6.17-6.05 (m, 1H), 4.37(s, 2H), 2.31 (s, 3H). MS: (ES) m/z calculated for C19H13ClF3N3O4 [M +H]+ 440.1, found 440.0.

1H NMR (400 MHz, DMSO-d6) δ 9.78 (s, 1H), 9.15 (s, 1H), 7.64- 7.57 (m,2H), 6.25 (d, J = 3.1 Hz, 1H), 6.05-6.02 (m, 1H), 4.85 (t, J = 5.4 Hz,1H), 4.53 (s, 2H), 3.65- 3.52 (m, 4H), 2.25 (s, 3H), 2.03- 1.78 (m, 2H),0.90 (t, J = 7.4 Hz, 3H). MS: (ES) m/z calculated for C22DH21ClN3O5 [M +Na]+ 467.1, found 467.1.

1H NMR (400 MHz, DMSO-d6) δ 9.75 (s, 1H), 9.15 (s, 1H), 7.63- 7.55 (m,2H), 6.25 (d, J = 3.1 Hz, 1H), 6.06-6.02 (m, 1H), 4.46 (s, 2H), 3.06 (s,3H), 2.25 (s, 3H), 2.00-1.80 (m,, 2H), 0.90 (t, J = 7.3 Hz, 3H). MS:(ES) m/z calculated for C21DH19ClN3O4 [M + H]+ 415.1, found 415.1.

1H NMR (400 MHz, DMSO-d6) δ 9.86 (s, 1H), 9.21 (s, 1H), 7.65- 7.55 (m,2H), 6.25 (d, J = 3.1 Hz, 1H), 6.04 (d, J = 3.1 Hz, 1H), 2.92 (s, 3H),2.24 (s, 3H), 2.00-1.80 (m, 2H), 1.55 (s, 6H), 0.90 (t, J = 7.3 Hz, 3H).MS: (ES) m/z calculated for C23DH23ClN3O4 [M + H]+ 443.1, found 443.1.

1H NMR (400 MHz, DMSO-d6) δ 9.77 (s, 1H), 9.13 (s, 1H), 7.60- 7.47 (m,2H), 7.18 (d, J = 7.3 Hz, 1H), 6.25 (d, J = 3.1 Hz, 1H), 6.06- 6.02 (m,1H), 4.45 (s, 2H), 3.04 (s, 3H), 2.25 (s, 3H), 2.00-1.80 (m, 2H), 0.91(t, J = 7.3 Hz, 3H). MS: (ES) m/z calculated for C21DH20N3O4 [M + H]+381.2, found 381.2.

1H NMR (400 MHz, DMSO-d6) δ 9.86 (s, 1H), 9.28-8.96 (m, 2H), 7.64-7.48(m, 2H), 6.30-6.16 (m, 1H), 6.07-5.95 (m, 1H), 2.23 (s, 3H), 1.98-1.78(m, 2H), 1.55 (s, 6H), 0.93-0.83 (m, 3H). MS: (ES) m/z calculated forC22DH21ClN3O4 [M + H]+ 429.1, found 429.1.

1H NMR (400 MHz, DMSO-d6) δ 9.76 (s, 1H), 9.16 (s, 1H), 8.93 (s, 1H),7.59 (s, 2H), 6.25 (d, J = 3.1 Hz, 1H), 6.05-6.02 (m, 1H), 4.36 (s, 2H),2.25 (s, 3H), 2.00- 1.80 (m, 2H), 0.90 (t, J = 7.3 Hz, 3H). MS: (ES) m/zcalculated for C20DH17ClN3O4 [M + H]+ 401.1, found 401.1.

1H NMR (400 MHz, DMSO-d6) δ 9.82 (s, 1H), 9.19 (d, J = 8.9 Hz, 1H), 9.05(s, 1H), 7.61-7.53 (m, 2H), 6.25 (d, J = 3.1 Hz, 1H), 6.05- 6.02 (m,1H), 5.20-5.10 (m, 1H), 2.24 (s, 3H), 2.01-1.80 (m, 2H), 1.55 (s, 6H),0.91 (t, J = 7.3 Hz, 3H). MS: (ES) m/z calculated for C22H22ClN3O4 [M +H]+ 428.1, found 428.1.

1H NMR (400 MHz, DMSO-d6) δ 9.77 (s, 1H), 9.13 (d, J = 9.0 Hz, 1H),7.59-7.48 (m, 2H), 7.19 (d, J = 7.2 Hz, 1H), 6.25 (d, J = 3.1 Hz, 1H),6.05-6.02 (m, 1H), 5.20-5.09 (m, 1H), 4.51 (s, 2H), 3.65 (t, J = 5.3 Hz,2H), 3.54 (t, J = 5.4 Hz, 2H), 3.24 (s, 3H), 2.25 (s, 3H), 2.00-1.81 (m,2H), 0.91 (t, J = 7.3 Hz, 3H). MS: (ES) m/z calculated for C23H25N3O5[M + H]+ 424.2, found 424.1.

1H NMR (400 MHz, DMSO-d6) δ 9.84 (s, 1H), 9.21 (d, J = 8.8 Hz, 1H), 9.06(s, 1H), 7.63-7.54 (m, 2H), 6.53-6.44 (m, 2H), 5.22- 5.12 (m, 1H),2.01-1.83 (m, 2H), 1.56 (s, 6H), 0.91 (t, J = 7.3 Hz, 3H). MS: (ES) m/zcalculated for C21H19Cl2N3O4 [M + H]+ 448.1, found 448.0.

1H NMR (400 MHz, DMSO-d6) δ 9.79 (s, 1H), 9.13 (d, J = 8.9 Hz, 1H),7.59-7.48 (m, 2H), 7.19 (d, J = 7.3 Hz, 1H), 6.25 (d, J = 3.1 Hz, 1H),6.05-6.02 (m, 1H), 5.19-5.09 (m, 1H), 4.86-4.82 (m, 1H), 4.54 (s, 2H),3.63-3.51 (m, 4H), 2.25 (s, 3H), 2.01-1.80 (m, 2H), 0.91 (t, J = 7.3 Hz,3H). MS: (ES) m/z calculated for C22H23N3O5 [M + Na]+ 432.2, found432.1.

1H NMR (400 MHz, DMSO-d6) δ 9.79 (s, 1H), 9.20 (d, J = 8.8 Hz, 1H), 8.95(s, 1H), 7.65-7.58 (m, 2H), 6.53 (d, J = 3.3 Hz, 1H), 6.48 (d, J = 3.3Hz, 1H), 5.23-5.13 (m, 1H), 4.38 (s, 2H), 2.04-1.82 (m, 2H), 0.93 (t, J= 7.3 Hz, 3H). MS: (ES) m/z calculated for C19H15Cl2N3O4 [M + H]+ 420.0,found 420.0.

1H NMR (400 MHz, DMSO-d6) δ 9.70 (s, 1H), 9.08 (d, J = 9.3 Hz, 1H), 8.90(s, 1H), 7.59-7.52 (m, 1H), 7.41 (t, J = 8.8 Hz, 1H), 6.25 (d, J = 3.1Hz, 1H), 6.06-6.02 (m, 1H), 5.19-5.09 (m, 1H), 4.45 (s, 2H), 2.25 (s,3H), 2.01- 1.80 (m, 2H), 0.91 (t, J = 7.3 Hz, 3H). MS: (ES) m/zcalculated for C20H18FN3O4 [M + H]+ 384.1, found 384.1.

1H NMR (400 MHz, DMSO-d6) δ 9.83 (s, 1H), 9.16 (d, J = 9.5 Hz, 1H), 8.92(s, 1H), 7.59 (d, J = 8.7 Hz, 1H), 7.53 (d, J = 8.7 Hz, 1H), 6.22 (d, J= 3.1 Hz, 1H), 6.05- 6.02 (m, 1H), 5.06-4.98 (m, 1H), 4.36 (s, 2H), 2.26(s, 3H), 2.24-2.13 (m, 1H), 0.96 (d, J = 6.7 Hz, 3H), 0.86 (d, J = 6.7Hz, 3H). MS: (ES) m/z calculated for C21H20ClN3O4 [M + H]+ 414.1, found414.1.

1H NMR (400 MHz, DMSO-d6) δ 9.74 (s, 1H), 9.18 (d, J = 8.6 Hz, 1H), 8.69(s, 1H), 7.62-7.47 (m, 2H), 7.17 (d, J = 7.3 Hz, 1H), 6.26 (d, J = 3.1Hz, 1H), 6.06-6.02 (m, 1H), 5.42-5.28 (m, 1H), 4.36 (s, 2H), 2.24 (s,3H), 1.55 (d, J = 6.9 Hz, 3H). MS: (ES) m/z calculated for C19H17N3O4[M + Na]+ 374.1, found 374.1.

1H NMR (400 MHz, DMSO-d6) δ 9.76 (s, 1H), 9.16 (d, J = 9.0 Hz, 1H), 8.92(s, 1H), 7.59 (s, 2H), 6.25 (d, J = 3.1 Hz, 1H), 6.05- 6.02 (m, 1H),5.19-5.09 (m, 1H), 4.36 (s, 2H), 2.25 (s, 3H), 2.01-1.80 (m, 2H), 0.91(t, J = 7.3 Hz, 3H). MS: (ES) m/z calculated for C20H18ClN3O4 [M + H]+400.1, found 400.0.

1H NMR (400 MHz, DMSO-d6) δ 9.84 (s, 1H), 9.15 (d, J = 9.5 Hz, 1H), 8.69(s, 1H), 7.54-7.46 (m, 2H), 7.18 (d, J = 6.7 Hz, 1H), 6.22 (d, J = 3.1Hz, 1H), 6.05-6.02 (m, 1H), 5.07-4.99 (m, 1H), 4.37 (s, 2H), 2.26 (s,3H), 2.23- 2.13 (m, 1H), 0.96 (d, J = 6.7 Hz, 3H), 0.86 (d, J = 6.7 Hz,3H). MS: (ES) m/z calculated for C21H21N3O4 [M + H]+ 380.1, found 380.1.

1H NMR (400 MHz, DMSO-d6) δ 9.77 (s, 1H), 9.13 (d, J = 8.9 Hz, 1H), 8.69(s, 1H), 7.58-7.48 (m, 2H), 7.17 (d, J = 7.1 Hz, 1H), 6.25 (d, J = 3.1Hz, 1H), 6.05-6.02 (m, 1H), 5.20-5.09 (m, 1H), 4.36 (s, 2H), 2.25 (s,3H), 2.02- 1.80 (m, 2H), 0.91 (t, J = 7.3 Hz, 3H). MS: (ES) m/zcalculated for C20H19N3O4 [M + H]+ 366.1, found 366.1.

Biological Example 1: Ligand Binding Assay for CXCR2 Activity

A ligand binding assay can be used to determine the ability of potentialCXCR2 antagonists to block the interaction between CXCR2 and any of itsligands. HEK-293 cells stably expressing CXCR2 or human neutrophilsexpressing CXCR2, are centrifuged and resuspended in assay buffer (20 mMHEPES pH 7.1, 140 mM NaCl, 1 mM CaCl₂, 5 mM MgCl₂, 0.1% sodium azide andwith 0.1% bovine serum albumin) to a concentration of 5×10⁵ cells/mL.Binding assays are set up as follows: Compounds for screening areserially diluted from a maximum of 20 μM, and 0.1 mL of cells containing5×10⁴ cells (for the HEK-293 cells) or 3×10⁴ cells (for the humanneutrophils) is added to each well containing compound. Then 0.1 mL of¹²⁵I labeled CXCL8 (obtained from PerkinElmer; Waltham, Mass.) dilutedin assay buffer to a final concentration of ˜50 pM, yielding ˜1 μCi perwell is added, and the plates are sealed and incubated for approximately3 hours at 25° C. on a shaker platform. Reactions are aspirated ontoGF/B glass filters pre-soaked in 0.3% polyethyleneimine (PEI) solution,on a vacuum cell harvester (Packard Instruments; Meriden, Conn.).Scintillation fluid (50 uL; Microscint 20, Packard Instruments) is addedto each well, the plates are sealed and radioactivity measured in a TopCount scintillation counter (Packard Instruments). Control wellscontaining either diluent only (for total counts) or 20 μM compound areused to calculate the percent of total inhibition for compound. Thecomputer program Prism from GraphPad, Inc. (San Diego, Ca) can be usedto calculate IC₅₀ values. IC₅₀ values are those concentrations requiredto reduce the binding of labeled CXCR8 to the receptor by 50%. Compoundsin FIG. 1 having an IC₅₀ value in the binding assay of less than 100 nMare labeled (+++); from 100-1000 nM are labeled (++); and less than orequal to 20 μM but above 1000 nM are labeled (+).

Biological Example 2: Migration/Chemotaxis Assay

A serum chemotaxis assay can be used to determine the efficacy ofpotential receptor antagonists at blocking the migration mediatedthrough chemokine receptors, such as CXCR2. This assay is routinelyperformed using the ChemoTX® microchamber system with a 5-μm pore-sizedpolycarbonate membrane. To begin such an assay, chemokine-receptorexpressing cells (in this case neutrophils isolated from human wholeblood) are collected by centrifugation at 400×g at room temperature,then suspended at 4 million/ml in human serum. The compound being testedis serially diluted from a maximum final concentration of 10M (or anequivalent volume of its solvent (DMSO)) and is then added to thecell/serum mixture. Separately, recombinant human CXCL5 (ENA-78) at itsEC₅₀ concentration (10 nM) is placed in the lower wells of the ChemoTX®plate. The 5-μm (pore size) polycarbonate membrane is placed onto theplate, and 20 μL of the cell/compound mixture is transferred onto eachwell of the membrane. The plates are incubated at 37° C. for 45 minutes,after which the polycarbonate membranes are removed and 5 μl of theDNA-intercalating agent CyQUANT (Invitrogen, Carlsbad, Calif.) is addedto the lower wells. The amount of fluorescence, corresponding to thenumber of migrated cells, is measured using a Spectrafluor Plus platereader (TECAN, San Jose, Calif.).

Particular embodiments of this invention are described herein, includingthe best mode known to the inventors for carrying out the invention.Upon reading the foregoing, description, variations of the disclosedembodiments may become apparent to individuals working in the art, andit is expected that those skilled artisans may employ such variations asappropriate. Accordingly, it is intended that the invention be practicedotherwise than as specifically described herein, and that the inventionincludes all modifications and equivalents of the subject matter recitedin the claims appended hereto as permitted by applicable law. Moreover,any combination of the above-described elements in all possiblevariations thereof is encompassed by the invention unless otherwiseindicated herein or otherwise clearly contradicted by context.

All publications, patent applications, accession numbers, and otherreferences cited in this specification are herein incorporated byreference as if each individual publication or patent application werespecifically and individually indicated to be incorporated by reference.

1.-10. (canceled)
 11. A pharmaceutical composition comprising a compoundhaving formula (Ia),

or a pharmaceutically acceptable salt thereof and a pharmaceuticallyacceptable excipient, wherein R¹ is selected from the group consistingof Cl and CH₃; R^(3b) is selected from the group consisting of H and D;R⁴ is a member selected from the group consisting of H and C₁₋₈ alkyl,wherein the C₁₋₈ alkyl is optionally substituted with —CONR^(a)R^(b),—OC(O)NR^(a)R^(b), —NR^(a)C(O)R^(b), —NR^(a)C(O)₂R, —NR^(a)R^(b), and—OR^(a), wherein each R^(a) and R^(b) is independently selected fromhydrogen, C₁₋₄ alkyl, C₁₋₄ hydroxyalkyl and C₁₋₄ haloalkyl, and R^(c) isselected from C₁₋₄ alkyl, C₁₋₄ hydroxyalkyl and C₁₋₄ haloalkyl; R^(5a)and R^(5b) are each members independently selected from the groupconsisting of H, F, Cl and CH₃; R^(6a) and R^(6b) are each membersindependently selected from the group consisting of H, C₁₋₄ alkyl, C₁₋₄hydroxyalkyl and C₁₋₄ haloalkyl; or optionally R^(6a) and R^(6b) aretaken together to form oxo (═O); or any salts, solvates, hydrates,N-oxides or rotamers thereof.
 12. The pharmaceutical composition ofclaim 11, or a pharmaceutically acceptable salt thereof, wherein R^(3b)is H; R⁴ is H or CH₃; R^(5a) is H, F or Cl; R^(5b) is H, F, Cl; R^(6a)and R^(6b) are independently selected from the group consisting of H andCH₃, or are taken together to form oxo (═O).
 13. The pharmaceuticalcomposition of claim 11, or a pharmaceutically acceptable salt thereof,wherein R^(3b) is D; R⁴ is H or CH₃; R^(5a) is H, F or Cl; R^(5b) is H,F, Cl; R^(6a) and R^(6b) are independently selected from the groupconsisting of H and CH₃, or are taken together to form oxo (═O). 14.-18.(canceled)
 19. The pharmaceutical composition of claim 11, furthercomprising one or more additional therapeutic agents.
 20. Thepharmaceutical composition of claim 19, wherein the one or moreadditional therapeutic agent is selected from the group consisting of acytotoxic chemotherapy, anti-cancer or anti-tumor vaccines,anti-immunocytokine therapies, immunocytokine therapies, chimericantigen receptor (CAR) T cell receptors, gene transfer therapy, andcheckpoint inhibitors.
 21. The pharmaceutical composition of claim 19,wherein the one or more additional therapeutic agent is selected fromthe group consisting of: drugs that block the activity of CTLA-4(CD152), PD-1 (CD279), PDL-1 (CD274), TIM-3, LAG-3 (CD223), VISTA, KIR,NKG2A, BTLA, PD-1H, TIGIT, CD96, 4-1BB (CD137), 4-1BBL (CD137L), GARP,CSF-1R, A2AR, CD73, CD47, tryptophan 2,3-dioxygenase (TDO) orindoleamine 2,3 dioxygenase (IDO), and agonists of OX40, GITR, 4-1BB,ICOS, STING or CD40.
 22. (canceled)
 23. (canceled)
 24. A method oftreating a disease or disorder selected from the group consisting ofpsoriasis, rheumatoid arthritis, radiation induced fibrotic lungdisease, autoimmune bullous dermatosis (AIBD), chronic obstructivepulmonary disease, and ozone-induced airway inflammation in a subject inneed thereof, said method comprising administering an effective amountof a pharmaceutical composition of claim 11 to said subject.
 25. Amethod of treating a cancer selected from the group consisting ofrhabdomyocarcoma, Lewis lung carcinoma (LLC), non-small cell lungcancer, esophageal squamous cell carcinoma, esophageal adenocarcinoma,renal cell carcinoma (RCC), colorectal cancer (CRC), acute myeloidleukemia (AML), breast cancer, gastric cancer, prostatic small cellneuroendocrine carcinoma (SCNC), liver cancer, glioblastoma, livercancer, oral squamous cell carcinoma, head and neck squamous cellcarcinoma, pancreatic cancer, thyroid papillary cancer, intrahepaticcholangiocellular carcinoma, hepatocellular carcinoma, bone cancer, andnasopharyngeal carcinoma in a subject in need thereof, said methodcomprising administering an effective amount of a pharmaceuticalcomposition of claim 11 to said subject.
 26. A method of claim 25,wherein the pharmaceutical composition, is used to treat cancer alone orin combination with one or more other anti-cancer therapies.
 27. Amethod of claim 26, wherein the pharmaceutical composition, is used totreat cancer in combination with one or more of a cytotoxicchemotherapy, an anti-cancer vaccine, an anti-tumor vaccines, ananti-immunocytokine, an immunocytokine therapy, and a chimeric antigenreceptor (CAR) T cell receptors, gene transfer therapy.
 28. A method ofclaim 26, wherein the pharmaceutical composition, is used to treatcancer in combination with one or more checkpoint inhibitor.
 29. Themethod of claim 26 wherein the one or more other anti-cancer therapy isselected from the group consisting of: drugs that block the activity ofCTLA-4 (CD152), PD-1 (CD279), PDL-1 (CD274), TIM-3, LAG-3 (CD223),VISTA, KIR, NKG2A, BTLA, PD-1H, TIGIT, CD96, 4-1BB (CD137), 4-1BBL(CD137L), GARP, CSF-1R, A2AR, CD73, CD47, tryptophan 2,3-dioxygenase(TDO) or indoleamine 2,3 dioxygenase (IDO), and agonists of OX40, GITR,4-1BB, ICOS, STING or CD40.
 30. The pharmaceutical composition of claim11, wherein the compound of formula (Ia) is selected from the groupconsisting of

or a pharmaceutically acceptable salt thereof.
 31. The pharmaceuticalcomposition of claim 11, wherein the compound of formula (Ia) isselected from the group consisting of


32. The pharmaceutical composition of claim 11, wherein the compound offormula (Ia) is selected from the group consisting of